From 12ae2c43318447f2cff8d84304f2991bc00d5df4 Mon Sep 17 00:00:00 2001 From: Joachim Wolff Date: Tue, 1 Oct 2024 16:27:33 +0200 Subject: [PATCH 1/6] New micro-c build of interaction matrix --- hicexplorer/hicBuildMatrix.py | 1381 +--------------- hicexplorer/hicBuildMatrixMicroC.py | 230 +++ hicexplorer/lib/buildMatrixMethods.py | 1405 +++++++++++++++++ .../test/general/test_hicBuildMatrixMicroC.py | 321 ++++ setup.py | 2 +- 5 files changed, 1967 insertions(+), 1372 deletions(-) create mode 100644 hicexplorer/hicBuildMatrixMicroC.py create mode 100644 hicexplorer/lib/buildMatrixMethods.py create mode 100644 hicexplorer/test/general/test_hicBuildMatrixMicroC.py diff --git a/hicexplorer/hicBuildMatrix.py b/hicexplorer/hicBuildMatrix.py index 409ce14c..2188fcfb 100644 --- a/hicexplorer/hicBuildMatrix.py +++ b/hicexplorer/hicBuildMatrix.py @@ -1,87 +1,19 @@ import argparse -import numpy as np -from scipy.sparse import coo_matrix, dia_matrix -import time -from os import unlink import os -from io import StringIO -import traceback + import warnings warnings.simplefilter(action="ignore", category=RuntimeWarning) warnings.simplefilter(action="ignore", category=PendingDeprecationWarning) -import pysam -from collections import OrderedDict - -from copy import deepcopy -from ctypes import Structure, c_uint, c_ushort -from multiprocessing import Process, Queue -from multiprocessing.sharedctypes import Array, RawArray - -from intervaltree import IntervalTree, Interval - -from Bio.Seq import Seq # own tools -from hicmatrix import HiCMatrix as hm -from hicexplorer.utilities import getUserRegion, genomicRegion +from hicexplorer.utilities import genomicRegion from hicexplorer._version import __version__ -import hicexplorer.hicPrepareQCreport as QC -from hicmatrix.lib import MatrixFileHandler - -from hicexplorer import hicMergeMatrixBins import logging log = logging.getLogger(__name__) - -class C_Interval(Structure): - """Struct to map a Interval form intervaltree as a multiprocessing.sharedctype""" - _fields_ = [("begin", c_uint), - ("end", c_uint), - ("data", c_uint)] - - -class C_Coverage(Structure): - """Struct to model the coverage as a multiprocessing.sharedctype""" - - _fields_ = [("begin", c_uint), - ("end", c_uint)] - - -class ReadPositionMatrix(object): - """A class to check for PCR duplicates. - A set storing all possible - start sites. Checks if read is already in the set. - """ - - def __init__(self): - """ - >>> rp = ReadPositionMatrix() - >>> rp.is_duplicated('1', 0, '2', 0) - False - >>> rp.is_duplicated('1', 0, '2', 0) - True - """ - - self.pos_matrix = set() - - def is_duplicated(self, chrom1, start1, chrom2, start2): - if chrom1 < chrom2: - id_string = "{}-{}".format(chrom1, chrom2) - else: - id_string = "{}-{}".format(chrom2, chrom1) - - if start1 < start2: - id_string += "-{}-{}".format(start1, start2) - else: - id_string += "-{}-{}".format(start2, start1) - - if id_string in self.pos_matrix: - return True - else: - self.pos_matrix.add(id_string) - return False +from hicexplorer.lib.buildMatrixMethods import * def parse_arguments(args=None): @@ -313,787 +245,6 @@ def parse_arguments(args=None): return parser -def intervalListToIntervalTree(interval_list): - r""" - given a dictionary containing tuples of chrom, start, end, - this is transformed to an interval trees. To each - interval an id is assigned, this id corresponds to the - position of the interval in the given array of tuples - and if needed can be used to identify - the index of a row/colum in the hic matrix. - - >>> bin_list = [('chrX', 0, 50000), ('chrX', 50000, 100000)] - >>> res = intervalListToIntervalTree(bin_list) - >>> sorted(res['chrX']) - [Interval(0, 50000, 0), Interval(50000, 100000, 1)] - """ - bin_int_tree = {} - - for intval_id, intval in enumerate(interval_list): - chrom, start, end = intval[0:3] - if chrom not in bin_int_tree: - bin_int_tree[chrom] = IntervalTree() - bin_int_tree[chrom].add(Interval(start, end, intval_id)) - - return bin_int_tree - - -def get_bins(bin_size, chrom_size, region=None): - r""" - Split the chromosomes into even sized bins - of length bin_size. - - Returns a list of tuples containing - ('chrom', start, end) - - >>> test = Tester() - >>> chrom_size = get_chrom_sizes(pysam.Samfile(test.bam_file_1)) - >>> sorted(get_bins(50000, chrom_size)) - [('contig-1', 0, 7125), ('contig-2', 0, 3345)] - >>> get_bins(50000, chrom_size, region='contig-1') - [('contig-1', 0, 7125)] - """ - bin_intvals = [] - start = 0 - if region: - chrom_size, start, _, _ = \ - getUserRegion(chrom_size, region) - - for chrom, size in chrom_size: - for interval in range(start, size, bin_size): - bin_intvals.append((chrom, interval, - min(size, interval + bin_size))) - return bin_intvals - - -def bed2interval_list(bed_file_handler, pChromosomeSize, pRegion): - r""" - reads a BED file and returns - a list of tuples containing - (chromosome name, start, end) - - >>> import tempfile, os - - Make a temporary BED file - >>> _file = tempfile.NamedTemporaryFile(delete=False) - >>> _file.close() - >>> file_tmp = open(_file.name, 'w') - >>> foo = file_tmp.write('chr1\t10\t20\tH1\t0\n') - >>> foo = file_tmp.write("chr1\t60\t70\tH2\t0\n") - >>> file_tmp.close() - >>> bed2interval_list(open(_file.name, 'r'), None, None) - [('chr1', 10, 20), ('chr1', 60, 70)] - >>> os.remove(_file.name) - """ - - if pRegion is not None: - chrom_size, start_region, end_region, _ = \ - getUserRegion(pChromosomeSize, pRegion) - - count = 0 - interval_list = [] - for line in bed_file_handler: - count += 1 - fields = line.strip().split() - try: - chrom, start, end = fields[0], int(fields[1]), int(fields[2]) - - except IndexError: - log.error("error reading BED file at line {}".format(count)) - - if pRegion is not None: - if chrom == chrom_size[0] and start_region <= start and end_region <= end: - interval_list.append((chrom, start, end)) - else: - interval_list.append((chrom, start, end)) - - return interval_list - - -def get_rf_bins(rf_cut_intervals, min_distance=200, max_distance=800): - r""" - returns a list of tuples containing a bin having the format: - ('chrom', start, end) - - The goal is to have bins containing each a restriction site close to - the center. Restriction sites that are less than 'min_distance' appart - are merged. To the left and right of the restriction site - 'max_distance' bp are added unless they clash with other bin. - Resulting bins are not immediately one after the other. - - The given cut sites list has to be ordered chr, and start site - - :param rf_cut_intervals: list of tuples containing the position of restriction fragment sites - :param min_distance: min distance between restriction fragment sites. - :param max_distance: max distance between restriction fragment and bin border. - :return: - - # two restriction sites of length 10 - >>> rf_cut_interval = [('chr1', 10, 20), ('chr1', 60, 70)] - - The following two bins are close together - and should be merged under min_distance = 20 - >>> rf_cut_interval.extend([('chr2', 20, 30), ('chr2', 40, 50), - ... ('chr2', 70, 80)]) - >>> get_rf_bins(rf_cut_interval, min_distance=10, max_distance=20) - [('chr1', 0, 40), ('chr1', 40, 90), ('chr2', 0, 60), ('chr2', 60, 100)] - """ - log.info("Minimum distance considered between " - "restriction sites is {}\nMax " - "distance: {}\n".format(min_distance, max_distance)) - - chrom, start, end = list(zip(*rf_cut_intervals)) - - rest_site_len = end[0] - start[0] - - # find sites that are less than min_distance apart - to_merge = np.flatnonzero(np.diff(start) - rest_site_len <= min_distance) - to_merge += 1 # + 1 to account for np.diff index handling - merge_idx = 0 - # add max_distance to both sides - start = np.array(start) - max_distance - end = np.array(end) + max_distance - new_start = [max(0, start[0])] - new_end = [] - new_chrom = [chrom[0]] - for idx in range(1, len(start)): - # identify end of chromosome - if chrom[idx] != chrom[idx - 1]: - new_start.append(max(0, start[idx])) - new_end.append(end[idx - 1]) - new_chrom.append(chrom[idx]) - merge_idx += 1 - continue - - if merge_idx < len(to_merge) and idx == to_merge[merge_idx]: - merge_idx += 1 - continue - - # identify overlapping bins - if chrom[idx] == chrom[idx - 1] and end[idx - 1] > start[idx]: - middle = start[idx] + int((end[idx - 1] - start[idx]) / 2) - new_start.append(middle) - new_end.append(middle) - else: - new_start.append(start[idx]) - new_end.append(end[idx - 1]) - new_chrom.append(chrom[idx]) - - new_end.append(end[-1]) - assert len(new_chrom) == len(new_start), "error" - assert len(new_end) == len(new_start), "error" - - intervals = zip(new_chrom, new_start, new_end) - intervals = [(_chrom, _start, _end) for _chrom, _start, - _end in intervals if _end - _start >= min_distance] - return intervals - - -def get_chrom_sizes(bam_handle): - """ - return the list of chromosome names and their - size from the bam file - The return value is a list of the form - [('chr1', 2343434), ('chr2', 43432432)] - - >>> test = Tester() - >>> get_chrom_sizes(pysam.Samfile(test.bam_file_1, 'rb')) - [('contig-1', 7125), ('contig-2', 3345)] - """ - - # in some cases there are repeated entries in - # the bam file. Thus, I first convert to dict, - # then to list. - list_chrom_sizes = OrderedDict(zip(bam_handle.references, - bam_handle.lengths)) - return list(list_chrom_sizes.items()) - - -def check_dangling_end(read, dangling_sequences): - """ - given a pysam read object, this function - checks if a forward read starts with - the dangling sequence or if a reverse - read ends with the dangling sequence. - """ - ds = dangling_sequences - # check if keys are existing, return false otherwise - if 'pat_forw' not in ds or 'pat_rev' not in ds: - return False - # skip forward read that stars with the restriction sequence - if not read.is_reverse and \ - read.seq.upper().startswith(ds['pat_forw']): - return True - - # skip reverse read that ends with the restriction sequence - if read.is_reverse and \ - read.seq.upper().endswith(ds['pat_rev']): - return True - - return False - - -def get_supplementary_alignment(read, pysam_obj): - """Checks if a read has a supplementary alignment - :param read pysam AlignedSegment - :param pysam_obj pysam file object - - :return pysam AlignedSegment of the supplementary aligment or None in case of no supplementary alignment - """ - - # the SA field contains a list of other alignments as a ';' delimited list in the format - # rname,pos,strand,CIGAR,mapQ,NM; - if read.has_tag('SA'): - # field always ends in ';' thus last element after split is always - # empty, hence [0:-1] - other_alignments = read.get_tag('SA').split(";")[0:-1] - supplementary_alignment = [] - for i in range(len(other_alignments)): - _sup = next(pysam_obj) - if _sup.qname == read.qname: - supplementary_alignment.append(_sup) - - return supplementary_alignment - - else: - return None - - -def get_correct_map(primary, supplement_list): - r""" - Decides which of the mappings, the primary or supplement, is correct. In the case of - long reads (eg. 150bp) the restriction enzyme site could split the read into two parts - but only the mapping corresponding to the start of the read should be considered as - the correct one. - - For example: - - Forward read: - - _ <- cut site - |======================|==========> - - - |----------------------| - correct mapping - - - reverse read: - - _ <- cut site - <======================|==========| - - - |----------| - correct mapping - - - :param primary: pysam AlignedSegment for primary mapping - :param supplement_list: list of pysam AlignedSegment for secondary mapping - - :return: pysam AlignedSegment that is mapped correctly - - Examples - -------- - >>> sam_file_name = "/tmp/test.sam" - >>> sam_file = open(sam_file_name, 'w') - >>> _ = sam_file.write('''@HD\tVN:1.0 - ... @SQ\tSN:chr1\tLN:1575\tAH:chr1:5000000-5010000 - ... read\t65\tchr1\t33\t20\t10S1D25M\t=\t200\t167\tAGCTTAGCTAGCTACCTATATCTTGGTCTTGGCCG\t<<<<<<<<<<<<<<<<<<<<<:<9/,&,22;;<<<\t - ... read\t2113\tchr1\t88\t30\t1S34M\t=\t500\t412\tACCTATATCTTGGCCTTGGCCGATGCGGCCTTGCA\t<<<<<;<<<<7;:<<<6;<<<<<<<<<<<<7<<<<\t - ... read\t2113\tchr1\t120\t30\t5S30M\t=\t500\t412\tACCTATATCTTGGCCTTGGCCGATGCGGCCTTGCA\t<<<<<;<<<<7;:<<<6;<<<<<<<<<<<<7<<<<''') - >>> sam_file.close() - >>> sam = pysam.Samfile(sam_file_name, "r") - >>> read_list = [read for read in sam] - >>> primary = read_list[0] - >>> secondary_list = read_list[1:] - >>> first_mapped = get_correct_map(primary, secondary_list) - - The first mapped read is the first secondary at position 88 (sam 1-based) = 87 (0-based) - >>> print(first_mapped.pos) - 87 - - """ - - for supplement in supplement_list: - assert primary.qname == supplement.qname, "ERROR, primary " \ - "and supplementary reads do not have the same id. The ids " \ - "are as follows\n{}\n{}".format(primary.qname, supplement.qname) - read_list = [primary] + supplement_list - first_mapped = [] - for idx, read in enumerate(read_list): - if read.is_reverse: - cigartuples = read.cigartuples[::-1] - else: - cigartuples = read.cigartuples[:] - - # For each read in read_list, calculate the position of the first match (operation M in CIGAR string) in the read sequence. - # The calculation is done by adding up the lengths of all the operations until the first match. - # CIGAR string is a list of tuples of (operation, length). Match is stored as CMATCH. - match_sum = 0 - for op, count in cigartuples: - if op == pysam.CMATCH: - break - match_sum += count - - first_mapped.append(match_sum) - # find which read has a cigar string that maps first than any of the - # others. - idx_min = first_mapped.index(min(first_mapped)) - - return read_list[idx_min] - - -def enlarge_bins(bin_intervals, chrom_sizes): - r""" - takes a list of consecutive but not - one after the other bin intervals - and joins them such that the - end and start of consecutive bins - is the same. - - >>> chrom_sizes = [('chr1', 100), ('chr2', 100)] - >>> bin_intervals = [('chr1', 10, 30), ('chr1', 50, 80), - ... ('chr2', 10, 60), ('chr2', 60, 90)] - >>> enlarge_bins(bin_intervals, chrom_sizes) - [('chr1', 0, 40), ('chr1', 40, 100), ('chr2', 0, 60), ('chr2', 60, 100)] - """ - # enlarge remaining bins - chr_start = True - chrom_sizes_dict = dict(chrom_sizes) - for idx in range(len(bin_intervals) - 1): - chrom, start, end = bin_intervals[idx] - chrom_next, start_next, end_next = bin_intervals[idx + 1] - if chr_start is True: - start = 0 - chr_start = False - if chrom == chrom_next and \ - end != start_next: - middle = start_next - int((start_next - end) / 2) - bin_intervals[idx] = (chrom, start, middle) - bin_intervals[idx + 1] = (chrom, middle, end_next) - if chrom != chrom_next: - bin_intervals[idx] = (chrom, start, chrom_sizes_dict[chrom]) - bin_intervals[idx + 1] = (chrom_next, 0, end_next) - - chrom, start, end = bin_intervals[-1] - bin_intervals[-1] = (chrom, start, chrom_sizes_dict[chrom]) - - return bin_intervals - - -def readBamFiles(pFileOneIterator, pFileTwoIterator, pNumberOfItemsPerBuffer, pSkipDuplicationCheck, pReadPosMatrix, pRefId2name, pMinMappingQuality): - """Read the two bam input files into n buffers each with pNumberOfItemsPerBuffer - with n = number of processes. The duplication check is handled here too.""" - buffer_mate1 = [] - buffer_mate2 = [] - duplicated_pairs = 0 - one_mate_unmapped = 0 - one_mate_not_unique = 0 - one_mate_low_quality = 0 - - all_data_read = False - j = 0 - iter_num = 0 - while j < pNumberOfItemsPerBuffer: - try: - mate1 = next(pFileOneIterator) - mate2 = next(pFileTwoIterator) - except StopIteration: - all_data_read = True - break - iter_num += 1 - - # skip 'not primary' alignments - while mate1.flag & 256 == 256: - try: - mate1 = next(pFileOneIterator) - except StopIteration: - all_data_read = True - break - while mate2.flag & 256 == 256: - try: - mate2 = next(pFileTwoIterator) - except StopIteration: - all_data_read = True - break - - assert mate1.qname == mate2.qname, "FATAL ERROR {} {} " \ - "Be sure that the sam files have the same read order " \ - "If using Bowtie2 or Hisat2 add " \ - "the --reorder option".format(mate1.qname, mate2.qname) - - # check for supplementary alignments - # (needs to be done before skipping any unmapped reads - # to keep the order of the two bam files in sync) - mate1_supplementary_list = get_supplementary_alignment( - mate1, pFileOneIterator) - mate2_supplementary_list = get_supplementary_alignment( - mate2, pFileTwoIterator) - - if mate1_supplementary_list: - mate1 = get_correct_map(mate1, mate1_supplementary_list) - - if mate2_supplementary_list: - mate2 = get_correct_map(mate2, mate2_supplementary_list) - - # skip if any of the reads is not mapped - if mate1.flag & 0x4 == 4 or mate2.flag & 0x4 == 4: - one_mate_unmapped += 1 - continue - - # skip if the read quality is low - if mate1.mapq < pMinMappingQuality or mate2.mapq < pMinMappingQuality: - # for bwa other way to test - # for multi-mapping reads is with a mapq = 0 - # the XS flag is not reliable. - if mate1.mapq == 0 & mate2.mapq == 0: - one_mate_not_unique += 1 - continue - - """ - # check if low quality is because of - # read being repetitive - # by reading the XS flag. - # The XS:i field is set by bowtie when a read is - # multi read and it contains the mapping score of the next - # best match - if 'XS' in dict(mate1.tags) or 'XS' in dict(mate2.tags): - one_mate_not_unique += 1 - continue - """ - - one_mate_low_quality += 1 - continue - - if pSkipDuplicationCheck is False: - if pReadPosMatrix.is_duplicated(pRefId2name[mate1.rname], - mate1.pos, - pRefId2name[mate2.rname], - mate2.pos): - duplicated_pairs += 1 - continue - buffer_mate1.append(mate1) - buffer_mate2.append(mate2) - j += 1 - - if all_data_read and len(buffer_mate1) != 0 and len(buffer_mate2) != 0: - return buffer_mate1, buffer_mate2, True, duplicated_pairs, one_mate_unmapped, one_mate_not_unique, one_mate_low_quality, iter_num - len(buffer_mate1) - if all_data_read and len(buffer_mate1) == 0 or len(buffer_mate2) == 0: - return None, None, True, duplicated_pairs, one_mate_unmapped, one_mate_not_unique, one_mate_low_quality, iter_num - len(buffer_mate1) - return buffer_mate1, buffer_mate2, False, duplicated_pairs, one_mate_unmapped, one_mate_not_unique, one_mate_low_quality, iter_num - len(buffer_mate1) - - -def process_data(pMateBuffer1, pMateBuffer2, pMinMappingQuality, - pKeepSelfCircles, pRestrictionSequence, pKeepSelfLigation, pMatrixSize, - pRfPositions, pRefId2name, - pDanglingSequences, pBinsize, pResultIndex, - pQueueOut, pTemplate, pOutputBamSet, pCounter, - pSharedBinIntvalTree, pDictBinIntervalTreeIndex, pCoverage, pCoverageIndex, - pOutputFileBufferDir, pRow, pCol, pData, - pMaxInsertSize, pQuickQCMode): - """ - This function computes for a given number of elements in pMateBuffer1 and pMaterBuffer2 a partial interaction matrix. - This function is used by multiple processes to speed up the computation. - All partial matrices are merged in the end into one interaction matrix. - - Parameters - ---------- - pMateBuffer1 : List of n reads of type 'pysam.libcalignedsegment.AlignedSegment' of sam input file 1 - pMateBuffer2 : List of n reads of type 'pysam.libcalignedsegment.AlignedSegment' of sam input file 2 - pMinMappingQuality : integer, minimum mapping quality of a read - pKeepSelfCircles : boolean, if self circles should be kept - pRestrictionSequence : List of String, the restriction sequence - pKeepSelfLigation : If self ligations should be removed - pMatrixSize : integer, the size of the interaction matrix - pRfPositions : intervalTree, only used if a restriction cut file and not a bin size was defined. - pRefId2name : Tuple, Maps a reference id to a name - pDanglingSequences : dict, dict of dangling sequences - pBinsize : integer, the size of the bins - pResultIndex : integer, number of processs, range(0, threads). Is returned via the queue to have access to the right row, col and data array after the computation. - pQueueOut : multiprocessing.Queue, queue to return the computed counting variables: - one_mate_unmapped, one_mate_low_quality, one_mate_not_unique, dangling_end, self_circle, self_ligation, same_fragment, - mate_not_close_to_rf, count_inward, count_outward, count_left, count_right, inter_chromosomal, short_range, long_range, - pair_added, len(pMateBuffer1), pResultIndex, pCounter - pTemplate : The template for the output bam file - pOutputBamSet : If a output bam file should be written. Depending on the input parameter '--outBam' - pOutputName : String, Name of the partial bam file - pCounter : integer, value which is returned to the main process. The main process can than write a pCounter.bam_done file - to signal the background process, which is merging the partial bam files into one, that this dataset can be merged. - pSharedBinIntvalTree : multiprocessing.sharedctype.RawArray of C_Interval, stores the interval tree in a 1D-RawArray. - pDictBinIntervalTreeIndex : dict, stores the information at which index position a given interval starts and ends in the 1D-array 'pSharedBinIntvalTree' - pCoverage : multiprocessing.sharedctype.Array of c_uint, Stores the coverage in a 1D-Array - pCoverageIndex : multiprocessing.sharedctype.RawArray of C_Coverage, stores the information in the 1D-array 'pCoverage' - pOutputFileBufferDir : String, the directory where the partial output bam files are buffered. Default is '/dev/shm/' - pRow : multiprocessing.sharedctype.RawArray of c_uint, Stores the row index information. It is available for all processes and does not need to be copied. - pCol : multiprocessing.sharedctype.RawArray of c_uint, stores the column index information. It is available for all processes and does not need to be copied. - pData : multiprocessing.sharedctype.RawArray of c_ushort, stores a 1 for each row - column pair. It is available for all processes and does not need to be copied. - pMaxInsertSize : maximum illumina insert size - """ - try: - - one_mate_unmapped = 0 - one_mate_low_quality = 0 - one_mate_not_unique = 0 - dangling_end = {} - if pRestrictionSequence is not None: - for restrictionSequence in pRestrictionSequence: - dangling_end[restrictionSequence] = 0 - self_circle = 0 - self_ligation = 0 - same_fragment = 0 - mate_not_close_to_rf = 0 - - count_inward = 0 - count_outward = 0 - count_left = 0 - count_right = 0 - inter_chromosomal = 0 - short_range = 0 - long_range = 0 - - pair_added = 0 - - iter_num = 0 - hic_matrix = None - - out_bam_index_buffer = [] - - if pMateBuffer1 is None or pMateBuffer2 is None: - - pQueueOut.put([hic_matrix, [one_mate_unmapped, one_mate_low_quality, one_mate_not_unique, dangling_end, self_circle, self_ligation, same_fragment, - mate_not_close_to_rf, count_inward, count_outward, - count_left, count_right, inter_chromosomal, short_range, long_range, pair_added, iter_num, pResultIndex, out_bam_index_buffer]]) - return - - while iter_num < len(pMateBuffer1) and iter_num < len(pMateBuffer2): - mate1 = pMateBuffer1[iter_num] - mate2 = pMateBuffer2[iter_num] - iter_num += 1 - - # check if reads belong to a bin - # - # pDictBinInterval stores the start and end position for each chromsome in the array 'pSharedBinIntvalTree' - # To get to the right interval a binary search is used. - mate_bins = [] - mate_is_unasigned = False - for mate in [mate1, mate2]: - mate_ref = pRefId2name[mate.rname] - # find the middle genomic position of the read. This is used to - # find the bin it belongs to. - read_middle = mate.pos + int(mate.qlen / 2) - try: - start, end = pDictBinIntervalTreeIndex[mate_ref] - middle_pos = int((start + end) / 2) - mate_bin = None - while not start > end: - if pSharedBinIntvalTree[middle_pos].begin <= read_middle and read_middle <= pSharedBinIntvalTree[middle_pos].end: - mate_bin = pSharedBinIntvalTree[middle_pos] - mate_is_unasigned = False - break - elif pSharedBinIntvalTree[middle_pos].begin > read_middle: - end = middle_pos - 1 - middle_pos = int((start + end) / 2) - mate_is_unasigned = True - else: - start = middle_pos + 1 - middle_pos = int((start + end) / 2) - mate_is_unasigned = True - - except KeyError: - # for small contigs it can happen that they are not - # in the bin_intval_tree keys if no restriction site is found - # on the contig. - mate_is_unasigned = True - break - - # report no match case - if mate_bin is None: - mate_is_unasigned = True - break - mate_bin_id = mate_bin.data - mate_bins.append(mate_bin_id) - - # if a mate is unassigned, it means it is not close - # to a restriction site - if mate_is_unasigned is True: - mate_not_close_to_rf += 1 - continue - - # check if mates are in the same chromosome - if mate1.reference_id != mate2.reference_id: - orientation = 'diff_chromosome' - else: - # to identify 'inward' and 'outward' orientations - # the order or the mates in the genome has to be - # known. - if mate1.pos < mate2.pos: - first_mate = mate1 - second_mate = mate2 - else: - first_mate = mate2 - second_mate = mate1 - - """ - outward - <--------------- ----------------> - - inward - ---------------> <---------------- - - same-strand-right - ---------------> ----------------> - - same-strand-left - <--------------- <---------------- - """ - - if not first_mate.is_reverse and second_mate.is_reverse: - orientation = 'inward' - elif first_mate.is_reverse and not second_mate.is_reverse: - orientation = 'outward' - elif first_mate.is_reverse and second_mate.is_reverse: - orientation = 'same-strand-left' - else: - orientation = 'same-strand-right' - - # check self-circles - # self circles are defined as outward pairs that do not - # have a restriction sequence in between. The distance of < 25kb is - # used to only check close outward pairs as far apart pairs can not be self-circles - if abs(mate2.pos - mate1.pos) < 25000 and orientation == 'outward': - has_rf = [] - - if pRfPositions and pRestrictionSequence: - # check if in between the two mate - # ends the restriction fragment is found. - # check for multiple restriction sequences - for restrictionSequence in pRestrictionSequence: - # the interval used is: - # start of fragment + length of restriction sequence - # end of fragment - length of restriction sequence - # the restriction sequence length is subtracted - # such that only fragments internally containing - # the restriction site are identified - frag_start = min(mate1.pos, mate2.pos) + \ - len(restrictionSequence) - frag_end = max(mate1.pos + mate1.qlen, mate2.pos + mate2.qlen) - len(restrictionSequence) - mate_ref = pRefId2name[mate1.rname] - if mate_ref in pRfPositions: - has_rf.extend(sorted( - pRfPositions[mate_ref][frag_start: frag_end])) - - if len(has_rf) == 0: - self_circle += 1 - if not pKeepSelfCircles: - continue - if abs(mate2.pos - mate1.pos) < pMaxInsertSize and orientation == 'inward': - # check for dangling ends if the restriction sequence is known and if they look - # like 'same fragment' - if pRestrictionSequence: - if pDanglingSequences: - - # check for dangling ends in sequence. Stop check with first match - one_match = False - for restrictionSequence in pRestrictionSequence: - if check_dangling_end(mate1, pDanglingSequences[restrictionSequence]) or \ - check_dangling_end(mate2, pDanglingSequences[restrictionSequence]): - dangling_end[restrictionSequence] += 1 - one_match = True - break - if one_match: - continue - has_rf = [] - - if pRfPositions and pRestrictionSequence: - # check if in between the two mate - # ends the restriction fragment is found. - - # the interval used is: - # start of fragment + length of restriction sequence - # end of fragment - length of restriction sequence - # the restriction sequence length is subtracted - # such that only fragments internally containing - # the restriction site are identified - - for restrictionSequence in pRestrictionSequence: - frag_start = min(mate1.pos, mate2.pos) + \ - len(restrictionSequence) - frag_end = max(mate1.pos + mate1.qlen, mate2.pos + mate2.qlen) - len(restrictionSequence) - mate_ref = pRefId2name[mate1.rname] - if mate_ref in pRfPositions: - has_rf.extend(sorted( - pRfPositions[mate_ref][frag_start: frag_end])) - - # case when there is no restriction fragment site between the - # mates - if len(has_rf) == 0: - same_fragment += 1 - continue - - self_ligation += 1 - - if not pKeepSelfLigation: - # skip self ligations - continue - - # set insert size to save bam - mate1.isize = mate2.pos - mate1.pos - mate2.isize = mate1.pos - mate2.pos - - # if mate_bins, which is set in the previous section - # does not have size=2, it means that one - # of the exceptions happened - if len(mate_bins) != 2: - continue - - # count type of pair (distance, orientation) - if mate1.reference_id != mate2.reference_id: - inter_chromosomal += 1 - - elif abs(mate2.pos - mate1.pos) < 20000: - short_range += 1 - else: - long_range += 1 - - if orientation == 'inward': - count_inward += 1 - elif orientation == 'outward': - count_outward += 1 - elif orientation == 'same-strand-left': - count_left += 1 - elif orientation == 'same-strand-right': - count_right += 1 - - for mate in [mate1, mate2]: - # fill in coverage vector - vec_start = int(max(0, mate.pos - mate_bin.begin) / pBinsize) - length_coverage = pCoverageIndex[mate_bin_id].end - \ - pCoverageIndex[mate_bin_id].begin - vec_end = min(length_coverage, int( - vec_start + len(mate.seq) / pBinsize)) - coverage_index = pCoverageIndex[mate_bin_id].begin + vec_start - coverage_end = pCoverageIndex[mate_bin_id].begin + vec_end - for i in range(coverage_index, coverage_end, 1): - pCoverage[i] += 1 - - if not pQuickQCMode: - pRow[pair_added] = mate_bins[0] - pCol[pair_added] = mate_bins[1] - pData[pair_added] = np.uint8(1) - - pair_added += 1 - if pOutputBamSet: - - out_bam_index_buffer.append(iter_num - 1) - - pQueueOut.put([[one_mate_unmapped, one_mate_low_quality, one_mate_not_unique, dangling_end, self_circle, self_ligation, same_fragment, - mate_not_close_to_rf, count_inward, count_outward, - count_left, count_right, inter_chromosomal, short_range, long_range, pair_added, len(pMateBuffer1), pResultIndex, pCounter, out_bam_index_buffer]]) - except Exception as exp: - pQueueOut.put('Fail: ' + str(exp) + traceback.format_exc()) - return - return - - def main(args=None): """ Reads line by line two bam files that are not sorted. @@ -1110,525 +261,13 @@ def main(args=None): """ args = parse_arguments().parse_args(args) - # args.outFileName.name = args.outFileName.name.strip() - # log.debug('args.outFileName.name: {}'.format(args.outFileName.name.endswith('.h5'))) - if not args.outFileName.name.endswith('.h5') and not args.outFileName.name.endswith('.cool'): - if '.mcool' not in args.outFileName.name: - log.error('Please define the file extension. h5 and cool are supported, or the specializations of cool, mcool. Given input {}'.format(args.outFileName.name)) - exit(1) - # for backwards compatibility - if args.maxDistance is not None: - args.maxLibraryInsertSize = args.maxDistance - try: - QC.make_sure_path_exists(args.QCfolder) - except OSError: - exit("Can't open/create QC folder path: {}. Please check".format(args.QCfolder)) - - if args.threads < 2: - args.threads = 2 - warnings.warn( - "\nAt least two threads need to be defined. Setting --threads = 2!s\n") - - if args.danglingSequence and not args.restrictionSequence: - exit("\nIf --danglingSequence is set, --restrictionSequence needs to be set too.\n") - - log.info("reading {} and {} to build hic_matrix\n".format(args.samFiles[0].name, - args.samFiles[1].name)) - str1 = pysam.Samfile(args.samFiles[0].name, 'rb') - str2 = pysam.Samfile(args.samFiles[1].name, 'rb') - - args.samFiles[0].close() - args.samFiles[1].close() - if not args.doTestRun: - if args.outBam: - args.outBam.close() - out_bam_file = pysam.Samfile(args.outBam.name, 'wb', template=str1) - - if args.chromosomeSizes is None: - chrom_sizes = get_chrom_sizes(str1) - else: - chrom_sizes = OrderedDict() - with open(args.chromosomeSizes.name, 'r') as file: - file_ = True - while file_: - file_ = file.readline().strip() - if file_ != '': - line_split = file_.split('\t') - chrom_sizes[line_split[0]] = int(line_split[1]) - chrom_sizes = list(chrom_sizes.items()) - - # log.debug('chrom_sizes {}'.format(chrom_sizes)) - read_pos_matrix = ReadPositionMatrix() - - rf_interval = [] - for restrictionCutFile in args.restrictionCutFile: - rf_interval.extend(bed2interval_list(restrictionCutFile, chrom_sizes, args.region)) - - rf_positions = intervalListToIntervalTree(rf_interval) - log.debug('rf_positions {}'.format(rf_positions.keys())) - if args.binSize: - bin_intervals = get_bins(args.binSize[0], chrom_sizes, args.region) - else: - bin_intervals = get_rf_bins(rf_interval, - min_distance=args.minDistance, - max_distance=args.maxLibraryInsertSize) - - matrix_size = len(bin_intervals) - bin_intval_tree = intervalListToIntervalTree(bin_intervals) - ref_id2name = str1.references - - # build c_type shared memory for the interval tree - shared_array_list = [] - index_dict = {} - end = -1 - for i, seq in enumerate(bin_intval_tree): - start = end + 1 - interval_list = [] - for interval in bin_intval_tree[seq]: - interval_list.append((interval.begin, interval.end, interval.data)) - end = start + len(bin_intval_tree[seq]) - 1 - index_dict[seq] = (start, end) - interval_list = sorted(interval_list) - shared_array_list.extend(interval_list) - shared_build_intval_tree = RawArray(C_Interval, shared_array_list) - bin_intval_tree = None - dangling_sequences = {} - if args.danglingSequence: - # build a list of dangling sequences - if args.restrictionSequence is not None: - for i in range(0, len(args.restrictionSequence)): - args.restrictionSequence[i] = args.restrictionSequence[i].upper() - args.danglingSequence[i] = args.danglingSequence[i].upper() - dangling_sequences[args.restrictionSequence[i]] = {} - dangling_sequences[args.restrictionSequence[i]]['pat_forw'] = args.danglingSequence[i] - dangling_sequences[args.restrictionSequence[i]]['pat_rev'] = str( - Seq(args.danglingSequence[i]).reverse_complement()) - - log.info("dangling sequences to check " - "are {}\n".format(dangling_sequences)) - - # initialize coverage vectors that - # save the number of reads that overlap - # a bin. - # To save memory, coverage is not measured by bp - # but by bins of length 10bp - binsize = 10 - number_of_elements_coverage = 0 - start_pos_coverage = [] - end_pos_coverage = [] - - for chrom, start, end in bin_intervals: - start_pos_coverage.append(number_of_elements_coverage) - - number_of_elements_coverage += (end - start) // binsize - end_pos_coverage.append(number_of_elements_coverage - 1) - pos_coverage = RawArray(C_Coverage, list(zip( - start_pos_coverage, end_pos_coverage))) - start_pos_coverage = None - end_pos_coverage = None - coverage = Array(c_uint, [0] * number_of_elements_coverage) - - # define global shared ctypes arrays for row, col and data - args.threads = args.threads - 1 - row = [None] * args.threads - col = [None] * args.threads - data = [None] * args.threads - for i in range(args.threads): - row[i] = RawArray(c_uint, args.inputBufferSize) - col[i] = RawArray(c_uint, args.inputBufferSize) - data[i] = RawArray(c_ushort, args.inputBufferSize) - - start_time = time.time() - - iter_num = 0 - # pair_added = 0 - # hic_matrix = None - - one_mate_unmapped = 0 - one_mate_low_quality = 0 - one_mate_not_unique = 0 - dangling_end = {} - if args.restrictionSequence is not None: - for restrictionSequence in args.restrictionSequence: - dangling_end[restrictionSequence] = 0 - - self_circle = 0 - self_ligation = 0 - same_fragment = 0 - mate_not_close_to_rf = 0 - duplicated_pairs = 0 - - count_inward = 0 - count_outward = 0 - count_left = 0 - count_right = 0 - inter_chromosomal = 0 - short_range = 0 - long_range = 0 - - pair_added = 0 - - # input buffer for bam files - buffer_workers1 = [None] * args.threads - buffer_workers2 = [None] * args.threads - - # output buffer to write bam with mate1 and mate2 pairs - process = [None] * args.threads - all_data_processed = False - hic_matrix = coo_matrix((matrix_size, matrix_size), dtype='uint32') - queue = [None] * args.threads - - all_threads_done = False - thread_done = [False] * args.threads - count_output = 0 - count_call_of_read_input = 0 - computed_pairs = 0 - - if args.doTestRun: - args.inputBufferSize = args.doTestRunLines - fail_flag = False - fail_message = '' - while not all_data_processed or not all_threads_done: - - for i in range(args.threads): - if queue[i] is None and not all_data_processed: - count_call_of_read_input += 1 - - buffer_workers1[i], buffer_workers2[i], all_data_processed, \ - duplicated_pairs_, one_mate_unmapped_, one_mate_not_unique_, \ - one_mate_low_quality_, iter_num_ = readBamFiles(pFileOneIterator=str1, - pFileTwoIterator=str2, - pNumberOfItemsPerBuffer=args.inputBufferSize, - pSkipDuplicationCheck=args.skipDuplicationCheck, - pReadPosMatrix=read_pos_matrix, - pRefId2name=ref_id2name, - pMinMappingQuality=args.minMappingQuality - ) - duplicated_pairs += duplicated_pairs_ - one_mate_unmapped += one_mate_unmapped_ - one_mate_not_unique += one_mate_not_unique_ - one_mate_low_quality += one_mate_low_quality_ - iter_num += iter_num_ - queue[i] = Queue() - thread_done[i] = False - computed_pairs += len(buffer_workers1[i]) - # create process to compute hic matrix for this buffer - process[i] = Process(target=process_data, kwargs=dict( - pMateBuffer1=buffer_workers1[i], - pMateBuffer2=buffer_workers2[i], - pMinMappingQuality=args.minMappingQuality, - pKeepSelfCircles=args.keepSelfCircles, - pRestrictionSequence=args.restrictionSequence, - pKeepSelfLigation=args.keepSelfLigation, - pMatrixSize=matrix_size, - pRfPositions=rf_positions, - pRefId2name=ref_id2name, - pDanglingSequences=dangling_sequences, - pBinsize=binsize, - pResultIndex=i, - pQueueOut=queue[i], - pTemplate=str1, - pOutputBamSet=args.outBam, - pCounter=count_output, - pSharedBinIntvalTree=shared_build_intval_tree, - pDictBinIntervalTreeIndex=index_dict, - pCoverage=coverage, - pCoverageIndex=pos_coverage, - pOutputFileBufferDir="", - pRow=row[i], - pCol=col[i], - pData=data[i], - pMaxInsertSize=args.maxLibraryInsertSize, - pQuickQCMode=args.doTestRun - )) - process[i].start() - count_output += 1 - - elif queue[i] is not None and not queue[i].empty(): - result = queue[i].get() - if 'Fail:' in result: - fail_flag = True - fail_message = result[6:] - else: - if result[0] is not None: - elements = result[0][15] - if hic_matrix is None: - hic_matrix = coo_matrix( - (data[i][:elements], (row[i][:elements], col[i][:elements])), shape=(matrix_size, matrix_size)) - else: - hic_matrix += coo_matrix( - (data[i][:elements], (row[i][:elements], col[i][:elements])), shape=(matrix_size, matrix_size)) - - for sequence in result[0][3]: - dangling_end[sequence] += result[0][3][sequence] - self_circle += result[0][4] - self_ligation += result[0][5] - same_fragment += result[0][6] - mate_not_close_to_rf += result[0][7] - - count_inward += result[0][8] - count_outward += result[0][9] - count_left += result[0][10] - count_right += result[0][11] - inter_chromosomal += result[0][12] - short_range += result[0][13] - long_range += result[0][14] - - pair_added += result[0][15] - iter_num += result[0][16] - - for bam_index in result[0][19]: - mate1 = buffer_workers1[i][bam_index] - mate2 = buffer_workers2[i][bam_index] - - mate1.flag |= 0x1 - mate2.flag |= 0x1 - - # set one read as the first in pair and the - # other as second - mate1.flag |= 0x40 - mate2.flag |= 0x80 - - # set chrom of mate - mate1.mrnm = mate2.rname - mate2.mrnm = mate1.rname - - # set position of mate - mate1.mpos = mate2.pos - mate2.mpos = mate1.pos - - out_bam_file.write(mate1) - out_bam_file.write(mate2) - - buffer_workers1[i] = None - buffer_workers2[i] = None - queue[i] = None - process[i].join() - process[i].terminate() - process[i] = None - thread_done[i] = True - - # caused by the architecture I try to display this output - # information after +-1e5 of 1e6 reads. - if iter_num % 1e6 < 100000: - elapsed_time = time.time() - start_time - log.info("processing {} lines took {:.2f} " - "secs ({:.1f} lines per " - "second)\n".format(iter_num, - elapsed_time, - iter_num / elapsed_time)) - log.info("{} ({:.2f}%) valid pairs added to matrix" - "\n".format(pair_added, float(100 * pair_added) / iter_num)) - if args.doTestRun and iter_num > args.doTestRunLines: - log.debug( - "\n## *WARNING*. Early exit because of --doTestRun parameter ##\n\n") - all_data_processed = True - thread_done[i] = True - break - elif all_data_processed and queue[i] is None: - thread_done[i] = True - else: - time.sleep(1) - - if all_data_processed: - all_threads_done = True - for thread in thread_done: - if not thread: - all_threads_done = False - if fail_flag: - log.error(fail_message) - exit(1) - else: - log.debug('Parallel stuff done') - if not args.doTestRun: - # the resulting matrix is only filled unevenly with some pairs - # int the upper triangle and others in the lower triangle. To construct - # the definite matrix I add the values from the upper and lower triangles - # and subtract the diagonal to avoid double counting it. - # The resulting matrix is symmetric. - if args.outBam: - out_bam_file.close() - - dia = dia_matrix(([hic_matrix.diagonal()], [0]), - shape=hic_matrix.shape) - hic_matrix = hic_matrix + hic_matrix.T - dia - # extend bins such that they are next to each other - bin_intervals = enlarge_bins(bin_intervals[:], chrom_sizes) - # compute max bin coverage - bin_max = [] - - for cover in pos_coverage: - max_element = 0 - for i in range(cover.begin, cover.end, 1): - if coverage[i] > max_element: - max_element = coverage[i] - if max_element == 0: - bin_max.append(np.nan) - else: - bin_max.append(max_element) - - chr_name_list, start_list, end_list = list(zip(*bin_intervals)) - bin_intervals = list(zip(chr_name_list, start_list, end_list, bin_max)) - hic_ma = hm.hiCMatrix() - hic_ma.setMatrix(hic_matrix, cut_intervals=bin_intervals) - - """ - if args.restrictionCutFile: - # load the matrix to mask those - # bins that most likely didn't - # have a restriction site that was cutted - - # reload the matrix as a Hi-C matrix object - hic_matrix = hm.hiCMatrix(args.outFileName.name) - - hic_matrix.maskBins(get_poor_bins(bin_max)) - hic_matrix.save(args.outFileName.name) - """ - if not args.keepSelfLigation: - msg = " (removed)" - else: - msg = " (not removed)" - - mappable_unique_high_quality_pairs = iter_num - \ - (one_mate_unmapped + one_mate_low_quality + one_mate_not_unique) - - intermediate_qc_log = StringIO() - - intermediate_qc_log.write(""" -File\t{}\t\t -Sequenced reads\t{}\t\t -Min rest. site distance\t{}\t\t -Max library insert size\t{}\t\t - -""".format(args.outFileName.name, iter_num, args.minDistance, args.maxLibraryInsertSize)) - - intermediate_qc_log.write( - "#\tcount\t(percentage w.r.t. total sequenced reads)\n") - - intermediate_qc_log.write("Pairs mappable, unique and high quality\t{}\t({:.2f})\n". - format(mappable_unique_high_quality_pairs, - 100 * float(mappable_unique_high_quality_pairs) / iter_num)) - - intermediate_qc_log.write("Hi-C contacts\t{}\t({:.2f})\n". - format(pair_added, 100 * float(pair_added) / iter_num)) - - intermediate_qc_log.write("One mate unmapped\t{}\t({:.2f})\n". - format(one_mate_unmapped, 100 * float(one_mate_unmapped) / iter_num)) - - intermediate_qc_log.write("One mate not unique\t{}\t({:.2f})\n". - format(one_mate_not_unique, 100 * float(one_mate_not_unique) / iter_num)) - - intermediate_qc_log.write("Low mapping quality\t{}\t({:.2f})\n". - format(one_mate_low_quality, 100 * float(one_mate_low_quality) / iter_num)) - - intermediate_qc_log.write( - "\n#\tcount\t(percentage w.r.t. mappable, unique and high quality pairs)\n") - - if len(dangling_end) > 0: - log.debug('dangling_sequences {}'.format(dangling_sequences.keys())) - log.debug('dangling_end {}'.format(dangling_end.keys())) - # log.debug('dangling_end {}'.format(res.keys())) - - for key in dangling_end: - # dangling_sequences[args.restrictionSequence[i]]['pat_forw'] - intermediate_qc_log.write("dangling end {} (restriction sequence {})\t{}\t({:.2f})\n". - format(dangling_sequences[key]['pat_forw'], key, dangling_end[key], 100 * float(dangling_end[key]) / mappable_unique_high_quality_pairs)) - else: - intermediate_qc_log.write("dangling end\t{}\t({:.2f})\n". - format(0, 100 * float(0) / mappable_unique_high_quality_pairs)) - intermediate_qc_log.write("self ligation{}\t{}\t({:.2f})\n". - format(msg, self_ligation, 100 * float(self_ligation) / mappable_unique_high_quality_pairs)) - - intermediate_qc_log.write("One mate not close to rest site\t{}\t({:.2f})\n". - format(mate_not_close_to_rf, 100 * float(mate_not_close_to_rf) / mappable_unique_high_quality_pairs)) - - intermediate_qc_log.write("same fragment\t{}\t({:.2f})\n". - format(same_fragment, 100 * float(same_fragment) / mappable_unique_high_quality_pairs)) - - intermediate_qc_log.write("self circle\t{}\t({:.2f})\n". - format(self_circle, 100 * float(self_circle) / mappable_unique_high_quality_pairs)) - - intermediate_qc_log.write("duplicated pairs\t{}\t({:.2f})\n". - format(duplicated_pairs, 100 * float(duplicated_pairs) / mappable_unique_high_quality_pairs)) - - if pair_added > 0: - intermediate_qc_log.write( - "\n#\tcount\t(percentage w.r.t. total valid pairs used)\n") - intermediate_qc_log.write("inter chromosomal\t{}\t({:.2f})\n". - format(inter_chromosomal, 100 * float(inter_chromosomal) / pair_added)) - - intermediate_qc_log.write("Intra short range (< 20kb)\t{}\t({:.2f})\n". - format(short_range, 100 * float(short_range) / pair_added)) - - intermediate_qc_log.write("Intra long range (>= 20kb)\t{}\t({:.2f})\n". - format(long_range, 100 * float(long_range) / pair_added)) - - intermediate_qc_log.write("Read pair type: inward pairs\t{}\t({:.2f})\n". - format(count_inward, 100 * float(count_inward) / pair_added)) - - intermediate_qc_log.write("Read pair type: outward pairs\t{}\t({:.2f})\n". - format(count_outward, 100 * float(count_outward) / pair_added)) - - intermediate_qc_log.write("Read pair type: left pairs\t{}\t({:.2f})\n". - format(count_left, 100 * float(count_left) / pair_added)) - - intermediate_qc_log.write("Read pair type: right pairs\t{}\t({:.2f})\n". - format(count_right, 100 * float(count_right) / pair_added)) - - log_file_name = os.path.join(args.QCfolder, "QC.log") - log_file = open(log_file_name, 'w') - log_file.write(intermediate_qc_log.getvalue()) - log_file.close() - log.debug('log_file_name {}'.format(log_file_name)) - log.debug('args.QCfolder {}'.format(args.QCfolder)) - - QC.main("-l {} -o {}".format(log_file_name, args.QCfolder).split()) - - args.outFileName.close() - # removing the empty file. Otherwise the save method - # will say that the file already exists. - unlink(args.outFileName.name) - - hic_metadata = {} - hic_metadata['statistics'] = intermediate_qc_log.getvalue() - hic_metadata['matrix-generated-by'] = np.string_( - 'HiCExplorer-' + __version__) - hic_metadata['matrix-generated-by-url'] = np.string_( - 'https://github.com/deeptools/HiCExplorer') - if args.genomeAssembly: - hic_metadata['genome-assembly'] = np.string_(args.genomeAssembly) - - intermediate_qc_log.close() - if args.outFileName.name.endswith('.mcool') and args.binSize is not None and len(args.binSize) > 2: - - matrixFileHandlerOutput = MatrixFileHandler( - pFileType='cool', pHiCInfo=hic_metadata) - matrixFileHandlerOutput.set_matrix_variables(hic_ma.matrix, - hic_ma.cut_intervals, - hic_ma.nan_bins, - hic_ma.correction_factors, - hic_ma.distance_counts) - matrixFileHandlerOutput.save(args.outFileName.name + '::/resolutions/' + str( - args.binSize[0]), pSymmetric=True, pApplyCorrection=False) - - for resolution in args.binSize[1:]: - hic_matrix_to_merge = deepcopy(hic_ma) - _mergeFactor = int(resolution) // args.binSize[0] - merged_matrix = hicMergeMatrixBins.merge_bins( - hic_matrix_to_merge, _mergeFactor) - matrixFileHandlerOutput = MatrixFileHandler( - pFileType='cool', pAppend=True, pHiCInfo=hic_metadata) - matrixFileHandlerOutput.set_matrix_variables(merged_matrix.matrix, - merged_matrix.cut_intervals, - merged_matrix.nan_bins, - merged_matrix.correction_factors, - merged_matrix.distance_counts) - matrixFileHandlerOutput.save(args.outFileName.name + '::/resolutions/' + str( - resolution), pSymmetric=True, pApplyCorrection=False) - - else: - if not args.doTestRun: - hic_ma.save(args.outFileName.name, pHiCInfo=hic_metadata) + + createMatrix(pOutFileName=args.outFileName, pMaxDistance=args.maxDistance, pMaxLibraryInsertSize=args.maxLibraryInsertSize, pQCfolder=args.QCfolder, + pThreads=args.threads, pDanglingSequence=args.danglingSequence, pRestrictionSequence=args.restrictionSequence, pSamFiles=args.samFiles, + pDoTestRun=args.doTestRun, pOutBam=args.outBam, pChromosomeSizes=args.chromosomeSizes, pRestrictionCutFile=args.restrictionCutFile, + pRegion=args.region, pBinSize=args.binSize, pInputBufferSize=args.inputBufferSize, pMinDistance=args.minDistance, + pDoTestRunLines=args.doTestRunLines, pSkipDuplicationCheck=args.skipDuplicationCheck, pMinMappingQuality=args.minMappingQuality, + pKeepSelfCircles=args.keepSelfCircles, pKeepSelfLigation=args.keepSelfLigation, pGenomeAssembly=args.genomeAssembly) class Tester(object): diff --git a/hicexplorer/hicBuildMatrixMicroC.py b/hicexplorer/hicBuildMatrixMicroC.py new file mode 100644 index 00000000..1e8038aa --- /dev/null +++ b/hicexplorer/hicBuildMatrixMicroC.py @@ -0,0 +1,230 @@ +import argparse +import os + +import warnings +warnings.simplefilter(action="ignore", category=RuntimeWarning) +warnings.simplefilter(action="ignore", category=PendingDeprecationWarning) + + +# own tools +from hicexplorer.utilities import genomicRegion +from hicexplorer._version import __version__ + +import logging +log = logging.getLogger(__name__) + +from hicexplorer.lib.buildMatrixMethods import * + + +def parse_arguments(args=None): + + parser = argparse.ArgumentParser( + formatter_class=argparse.ArgumentDefaultsHelpFormatter, + add_help=False, + description=('Using an alignment from a program that supports ' + 'local alignment (eg. Bowtie2) where both ' + 'PE reads are mapped using the --local ' + 'option, this program reads such file and ' + 'creates a matrix of interactions.' + )) + + parserRequired = parser.add_argument_group('Required arguments') + + # define the arguments + parserRequired.add_argument('--samFiles', '-s', + help='The two PE alignment sam files to process', + metavar='two sam files', + nargs=2, + type=argparse.FileType('r'), + required=True) + + parserRequired.add_argument('--outFileName', '-o', + help='Output file name for the Hi-C matrix.', + metavar='FILENAME', + type=argparse.FileType('w'), + required=True) + + parserRequired.add_argument('--QCfolder', + help='Path of folder to save the quality control data for the matrix. The log files ' + 'produced this way can be loaded into `hicQC` in order to compare the quality of multiple ' + 'Hi-C libraries.', + metavar='FOLDER', + required=True) + + + parserOpt = parser.add_argument_group('Optional arguments') + + parserOpt.add_argument('--outBam', '-b', + help='Output bam file to process. Optional parameter. ' + 'A bam file containing all valid Hi-C reads can be created ' + 'using this option. This bam file could be useful to inspect ' + 'the distribution of valid Hi-C reads pairs or for other ' + 'downstream analyses, but is not used by any HiCExplorer tool. ' + 'Computation will be significantly longer if this option is set.', + metavar='bam file', + type=argparse.FileType('w'), + required=False) + + # group = parserOpt.add_mutually_exclusive_group(required=True) + + parserOpt.add_argument('--binSize', '-bs', + help='Size in bp for the bins. The bin size depends ' + 'on the depth of sequencing. Use a larger bin size for ' + 'libraries sequenced with lower depth. If not given, matrices of restriction site resolution will be built. ' + 'Optionally for mcool file format: Define multiple resolutions which are all a multiple of the first value. ' + ' Example: --binSize 10000 20000 50000 will create a mcool file formate containing the three defined resolutions.', + type=int, + nargs='+', + required=True) + + + + parserOpt.add_argument('--maxLibraryInsertSize', + help='The maximum library insert size defines different cut offs based on the maximum expected ' + 'library size. *This is not the average fragment size* but the higher end of the ' + 'the fragment size distribution (obtained using for example a Fragment Analyzer or a Bioanalyzer) ' + 'which usually is between 800 to 1500 bp. If this value is not known use the default value.' + ' The insert value is used to decide if two mates belong to the same fragment (by ' + 'checking if they are within this max insert size) and to decide if a mate is too far ' + 'away from the nearest restriction site' + ' (Default: %(default)s).', + type=int, + default=1000, + required=False) + + parserOpt.add_argument('--genomeAssembly', '-ga', + help='The genome the reads were mapped to. Used for metadata of cool file.') + + parserOpt.add_argument('--region', '-r', + help='Region of the genome to limit the operation to. ' + 'The format is chr:start-end. It is also possible to just ' + 'specify a chromosome, for example --region chr10', + metavar="CHR:START-END", + required=False, + type=genomicRegion + ) + + + parserOpt.add_argument('--keepSelfCircles', + help='If set, outward facing reads without any restriction fragment (self circles) are kept. ' + 'They will be counted and shown in the QC plots.', + required=False, + action='store_true' + ) + + parserOpt.add_argument('--minMappingQuality', + help='minimum mapping quality for reads to be accepted. ' + 'Because the restriction enzyme site could be located ' + 'on top of the read, this may reduce the ' + 'reported quality of the read. Thus, this parameter ' + 'may be adjusted if too many low quality ' + '(but otherwise perfectly valid Hi-C reads) are found. ' + 'A good strategy is to make a test run (using the --doTestRun), ' + 'then checking the results to see if too many low quality ' + 'reads are present and then using the bam file generated to ' + 'check if those low quality reads are caused by the read ' + 'not being mapped entirely' + ' (Default: %(default)s).', + required=False, + default=15, + type=int + ) + parserOpt.add_argument('--threads', + help='Number of threads. Using the python multiprocessing module. ' + 'One master process which is used to read the input file into the buffer and one process which is merging ' + 'the output bam files of the processes into one output bam file. All other threads do the actual computation. ' + 'Minimum value for the \'--thread\' parameter is 2. ' + 'The usage of 8 threads is optimal if you have an HDD. A higher number of threads is only ' + 'useful if you have a fast SSD. Have in mind that the performance of hicBuildMatrix is influenced by ' + 'the number of threads, the speed of your hard drive and the inputBufferSize. To clarify: the performance ' + 'with a higher thread number is not negative influenced but not positive too. With a slow HDD and a high number of ' + 'threads many threads will do nothing most of the time' + ' (Default: %(default)s).', + required=False, + default=4, + type=int + ) + parserOpt.add_argument('--inputBufferSize', + help='Size of the input buffer of each thread. 400,000 read pairs per input file per thread is the default value. ' + 'Reduce this value to decrease memory usage.', + required=False, + default=400000, + type=int + ) + parserOpt.add_argument('--doTestRun', + help='A test run is useful to test the quality ' + 'of a Hi-C experiment quickly. It works by ' + 'testing only 1,000,000 reads. This option ' + 'is useful to get an idea of quality control ' + 'values like inter-chromosomal interactions, ' + 'duplication rates etc.', + action='store_true' + ) + parserOpt.add_argument('--doTestRunLines', + help='Number of lines to consider for the qc test run' + ' (Default: %(default)s).', + required=False, + default=1000000, + type=int + ) + + parserOpt.add_argument('--skipDuplicationCheck', + help='Identification of duplicated read pairs is ' + 'memory consuming. Thus, in case of memory ' + 'errors this check can be skipped. However, ' + 'consider running a `--doTestRun` first to ' + 'get an estimation of the duplicated reads. ', + action='store_true' + ) + parserOpt.add_argument('--chromosomeSizes', '-cs', + help=('File with the chromosome sizes for your genome. A tab-delimited two column layout \"chr_name size\" is expected' + 'Usually the sizes can be determined from the SAM/BAM input files, however, ' + 'for cHi-C or scHi-C it can be that at the start or end no data is present. ' + 'Please consider that this option causes that only reads are considered which are on the listed chromosomes.' + 'Use this option to guarantee fixed sizes. An example file is available via UCSC: ' + 'http://hgdownload.soe.ucsc.edu/goldenPath/dm3/bigZips/dm3.chrom.sizes'), + type=argparse.FileType('r'), + metavar='txt file') + parserOpt.add_argument("--help", "-h", action="help", + help="show this help message and exit") + + parserOpt.add_argument('--version', action='version', + version='%(prog)s {}'.format(__version__)) + + return parser + + +def main(args=None): + """ + Reads line by line two bam files that are not sorted. + Each line in the two bam files should correspond + to the mapped position of the two ends of a Hi-C + fragment. + + Each mate pair is assessed to determine if it is + a valid Hi-C pair, in such case a matrix + reporting the counts of mates is constructed. + + A bam file containing the valid Hi-C reads + is also constructed. + """ + + args = parse_arguments().parse_args(args) + + createMatrix(pOutFileName=args.outFileName, pMaxDistance=None, pMaxLibraryInsertSize=args.maxLibraryInsertSize, pQCfolder=args.QCfolder, + pThreads=args.threads, pDanglingSequence=None, pRestrictionSequence=None, pSamFiles=args.samFiles, + pDoTestRun=args.doTestRun, pOutBam=args.outBam, pChromosomeSizes=args.chromosomeSizes, pRestrictionCutFile=None, + pRegion=args.region, pBinSize=args.binSize, pInputBufferSize=args.inputBufferSize, pMinDistance=None, + pDoTestRunLines=args.doTestRunLines, pSkipDuplicationCheck=args.skipDuplicationCheck, pMinMappingQuality=args.minMappingQuality, + pKeepSelfCircles=args.keepSelfCircles, pKeepSelfLigation=None, pGenomeAssembly=args.genomeAssembly) + + +class Tester(object): + def __init__(self): + hic_test_data_dir = os.environ.get('HIC_TEST_DATA_DIR', False) + if hic_test_data_dir: + self.root = hic_test_data_dir + else: + self.root = os.path.dirname( + os.path.abspath(__file__)) + "/test/test_data/" + self.bam_file_1 = os.path.join(self.root, "hic.bam") diff --git a/hicexplorer/lib/buildMatrixMethods.py b/hicexplorer/lib/buildMatrixMethods.py new file mode 100644 index 00000000..5f7e7628 --- /dev/null +++ b/hicexplorer/lib/buildMatrixMethods.py @@ -0,0 +1,1405 @@ +import logging +log = logging.getLogger(__name__) + +import argparse +import numpy as np +from scipy.sparse import coo_matrix, dia_matrix +import time +from os import unlink +import os +from io import StringIO +import traceback +import warnings +warnings.simplefilter(action="ignore", category=RuntimeWarning) +warnings.simplefilter(action="ignore", category=PendingDeprecationWarning) + +import pysam +from collections import OrderedDict + +from copy import deepcopy +from ctypes import Structure, c_uint, c_ushort +from multiprocessing import Process, Queue +from multiprocessing.sharedctypes import Array, RawArray + +from intervaltree import IntervalTree, Interval + +from Bio.Seq import Seq + +# own tools +from hicmatrix import HiCMatrix as hm +from hicexplorer.utilities import getUserRegion, genomicRegion +from hicexplorer._version import __version__ +import hicexplorer.hicPrepareQCreport as QC + +from hicmatrix.lib import MatrixFileHandler + +from hicexplorer import hicMergeMatrixBins +import logging +log = logging.getLogger(__name__) + +class C_Interval(Structure): + """Struct to map a Interval form intervaltree as a multiprocessing.sharedctype""" + _fields_ = [("begin", c_uint), + ("end", c_uint), + ("data", c_uint)] + + +class C_Coverage(Structure): + """Struct to model the coverage as a multiprocessing.sharedctype""" + + _fields_ = [("begin", c_uint), + ("end", c_uint)] + + +class ReadPositionMatrix(object): + """A class to check for PCR duplicates. + A set storing all possible + start sites. Checks if read is already in the set. + """ + + def __init__(self): + """ + >>> rp = ReadPositionMatrix() + >>> rp.is_duplicated('1', 0, '2', 0) + False + >>> rp.is_duplicated('1', 0, '2', 0) + True + """ + + self.pos_matrix = set() + + def is_duplicated(self, chrom1, start1, chrom2, start2): + if chrom1 < chrom2: + id_string = "{}-{}".format(chrom1, chrom2) + else: + id_string = "{}-{}".format(chrom2, chrom1) + + if start1 < start2: + id_string += "-{}-{}".format(start1, start2) + else: + id_string += "-{}-{}".format(start2, start1) + + if id_string in self.pos_matrix: + return True + else: + self.pos_matrix.add(id_string) + return False + +def intervalListToIntervalTree(interval_list): + r""" + given a dictionary containing tuples of chrom, start, end, + this is transformed to an interval trees. To each + interval an id is assigned, this id corresponds to the + position of the interval in the given array of tuples + and if needed can be used to identify + the index of a row/colum in the hic matrix. + + >>> bin_list = [('chrX', 0, 50000), ('chrX', 50000, 100000)] + >>> res = intervalListToIntervalTree(bin_list) + >>> sorted(res['chrX']) + [Interval(0, 50000, 0), Interval(50000, 100000, 1)] + """ + bin_int_tree = {} + + for intval_id, intval in enumerate(interval_list): + chrom, start, end = intval[0:3] + if chrom not in bin_int_tree: + bin_int_tree[chrom] = IntervalTree() + bin_int_tree[chrom].add(Interval(start, end, intval_id)) + + return bin_int_tree + + +def get_bins(bin_size, chrom_size, region=None): + r""" + Split the chromosomes into even sized bins + of length bin_size. + + Returns a list of tuples containing + ('chrom', start, end) + + >>> test = Tester() + >>> chrom_size = get_chrom_sizes(pysam.Samfile(test.bam_file_1)) + >>> sorted(get_bins(50000, chrom_size)) + [('contig-1', 0, 7125), ('contig-2', 0, 3345)] + >>> get_bins(50000, chrom_size, region='contig-1') + [('contig-1', 0, 7125)] + """ + bin_intvals = [] + start = 0 + if region: + chrom_size, start, _, _ = \ + getUserRegion(chrom_size, region) + + for chrom, size in chrom_size: + for interval in range(start, size, bin_size): + bin_intvals.append((chrom, interval, + min(size, interval + bin_size))) + return bin_intvals + + +def bed2interval_list(bed_file_handler, pChromosomeSize, pRegion): + r""" + reads a BED file and returns + a list of tuples containing + (chromosome name, start, end) + + >>> import tempfile, os + + Make a temporary BED file + >>> _file = tempfile.NamedTemporaryFile(delete=False) + >>> _file.close() + >>> file_tmp = open(_file.name, 'w') + >>> foo = file_tmp.write('chr1\t10\t20\tH1\t0\n') + >>> foo = file_tmp.write("chr1\t60\t70\tH2\t0\n") + >>> file_tmp.close() + >>> bed2interval_list(open(_file.name, 'r'), None, None) + [('chr1', 10, 20), ('chr1', 60, 70)] + >>> os.remove(_file.name) + """ + + if pRegion is not None: + chrom_size, start_region, end_region, _ = \ + getUserRegion(pChromosomeSize, pRegion) + + count = 0 + interval_list = [] + for line in bed_file_handler: + count += 1 + fields = line.strip().split() + try: + chrom, start, end = fields[0], int(fields[1]), int(fields[2]) + + except IndexError: + log.error("error reading BED file at line {}".format(count)) + + if pRegion is not None: + if chrom == chrom_size[0] and start_region <= start and end_region <= end: + interval_list.append((chrom, start, end)) + else: + interval_list.append((chrom, start, end)) + + return interval_list + + +def get_rf_bins(rf_cut_intervals, min_distance=200, max_distance=800): + r""" + returns a list of tuples containing a bin having the format: + ('chrom', start, end) + + The goal is to have bins containing each a restriction site close to + the center. Restriction sites that are less than 'min_distance' appart + are merged. To the left and right of the restriction site + 'max_distance' bp are added unless they clash with other bin. + Resulting bins are not immediately one after the other. + + The given cut sites list has to be ordered chr, and start site + + :param rf_cut_intervals: list of tuples containing the position of restriction fragment sites + :param min_distance: min distance between restriction fragment sites. + :param max_distance: max distance between restriction fragment and bin border. + :return: + + # two restriction sites of length 10 + >>> rf_cut_interval = [('chr1', 10, 20), ('chr1', 60, 70)] + + The following two bins are close together + and should be merged under min_distance = 20 + >>> rf_cut_interval.extend([('chr2', 20, 30), ('chr2', 40, 50), + ... ('chr2', 70, 80)]) + >>> get_rf_bins(rf_cut_interval, min_distance=10, max_distance=20) + [('chr1', 0, 40), ('chr1', 40, 90), ('chr2', 0, 60), ('chr2', 60, 100)] + """ + log.info("Minimum distance considered between " + "restriction sites is {}\nMax " + "distance: {}\n".format(min_distance, max_distance)) + + chrom, start, end = list(zip(*rf_cut_intervals)) + + rest_site_len = end[0] - start[0] + + # find sites that are less than min_distance apart + to_merge = np.flatnonzero(np.diff(start) - rest_site_len <= min_distance) + to_merge += 1 # + 1 to account for np.diff index handling + merge_idx = 0 + # add max_distance to both sides + start = np.array(start) - max_distance + end = np.array(end) + max_distance + new_start = [max(0, start[0])] + new_end = [] + new_chrom = [chrom[0]] + for idx in range(1, len(start)): + # identify end of chromosome + if chrom[idx] != chrom[idx - 1]: + new_start.append(max(0, start[idx])) + new_end.append(end[idx - 1]) + new_chrom.append(chrom[idx]) + merge_idx += 1 + continue + + if merge_idx < len(to_merge) and idx == to_merge[merge_idx]: + merge_idx += 1 + continue + + # identify overlapping bins + if chrom[idx] == chrom[idx - 1] and end[idx - 1] > start[idx]: + middle = start[idx] + int((end[idx - 1] - start[idx]) / 2) + new_start.append(middle) + new_end.append(middle) + else: + new_start.append(start[idx]) + new_end.append(end[idx - 1]) + new_chrom.append(chrom[idx]) + + new_end.append(end[-1]) + assert len(new_chrom) == len(new_start), "error" + assert len(new_end) == len(new_start), "error" + + intervals = zip(new_chrom, new_start, new_end) + intervals = [(_chrom, _start, _end) for _chrom, _start, + _end in intervals if _end - _start >= min_distance] + return intervals + + +def get_chrom_sizes(bam_handle): + """ + return the list of chromosome names and their + size from the bam file + The return value is a list of the form + [('chr1', 2343434), ('chr2', 43432432)] + + >>> test = Tester() + >>> get_chrom_sizes(pysam.Samfile(test.bam_file_1, 'rb')) + [('contig-1', 7125), ('contig-2', 3345)] + """ + + # in some cases there are repeated entries in + # the bam file. Thus, I first convert to dict, + # then to list. + list_chrom_sizes = OrderedDict(zip(bam_handle.references, + bam_handle.lengths)) + return list(list_chrom_sizes.items()) + + +def check_dangling_end(read, dangling_sequences): + """ + given a pysam read object, this function + checks if a forward read starts with + the dangling sequence or if a reverse + read ends with the dangling sequence. + """ + ds = dangling_sequences + # check if keys are existing, return false otherwise + if 'pat_forw' not in ds or 'pat_rev' not in ds: + return False + # skip forward read that stars with the restriction sequence + if not read.is_reverse and \ + read.seq.upper().startswith(ds['pat_forw']): + return True + + # skip reverse read that ends with the restriction sequence + if read.is_reverse and \ + read.seq.upper().endswith(ds['pat_rev']): + return True + + return False + + +def get_supplementary_alignment(read, pysam_obj): + """Checks if a read has a supplementary alignment + :param read pysam AlignedSegment + :param pysam_obj pysam file object + + :return pysam AlignedSegment of the supplementary aligment or None in case of no supplementary alignment + """ + + # the SA field contains a list of other alignments as a ';' delimited list in the format + # rname,pos,strand,CIGAR,mapQ,NM; + if read.has_tag('SA'): + # field always ends in ';' thus last element after split is always + # empty, hence [0:-1] + other_alignments = read.get_tag('SA').split(";")[0:-1] + supplementary_alignment = [] + for i in range(len(other_alignments)): + _sup = next(pysam_obj) + if _sup.qname == read.qname: + supplementary_alignment.append(_sup) + + return supplementary_alignment + + else: + return None + + +def get_correct_map(primary, supplement_list): + r""" + Decides which of the mappings, the primary or supplement, is correct. In the case of + long reads (eg. 150bp) the restriction enzyme site could split the read into two parts + but only the mapping corresponding to the start of the read should be considered as + the correct one. + + For example: + + Forward read: + + _ <- cut site + |======================|==========> + - + |----------------------| + correct mapping + + + reverse read: + + _ <- cut site + <======================|==========| + - + |----------| + correct mapping + + + :param primary: pysam AlignedSegment for primary mapping + :param supplement_list: list of pysam AlignedSegment for secondary mapping + + :return: pysam AlignedSegment that is mapped correctly + + Examples + -------- + >>> sam_file_name = "/tmp/test.sam" + >>> sam_file = open(sam_file_name, 'w') + >>> _ = sam_file.write('''@HD\tVN:1.0 + ... @SQ\tSN:chr1\tLN:1575\tAH:chr1:5000000-5010000 + ... read\t65\tchr1\t33\t20\t10S1D25M\t=\t200\t167\tAGCTTAGCTAGCTACCTATATCTTGGTCTTGGCCG\t<<<<<<<<<<<<<<<<<<<<<:<9/,&,22;;<<<\t + ... read\t2113\tchr1\t88\t30\t1S34M\t=\t500\t412\tACCTATATCTTGGCCTTGGCCGATGCGGCCTTGCA\t<<<<<;<<<<7;:<<<6;<<<<<<<<<<<<7<<<<\t + ... read\t2113\tchr1\t120\t30\t5S30M\t=\t500\t412\tACCTATATCTTGGCCTTGGCCGATGCGGCCTTGCA\t<<<<<;<<<<7;:<<<6;<<<<<<<<<<<<7<<<<''') + >>> sam_file.close() + >>> sam = pysam.Samfile(sam_file_name, "r") + >>> read_list = [read for read in sam] + >>> primary = read_list[0] + >>> secondary_list = read_list[1:] + >>> first_mapped = get_correct_map(primary, secondary_list) + + The first mapped read is the first secondary at position 88 (sam 1-based) = 87 (0-based) + >>> print(first_mapped.pos) + 87 + + """ + + for supplement in supplement_list: + assert primary.qname == supplement.qname, "ERROR, primary " \ + "and supplementary reads do not have the same id. The ids " \ + "are as follows\n{}\n{}".format(primary.qname, supplement.qname) + read_list = [primary] + supplement_list + first_mapped = [] + for idx, read in enumerate(read_list): + if read.is_reverse: + cigartuples = read.cigartuples[::-1] + else: + cigartuples = read.cigartuples[:] + + # For each read in read_list, calculate the position of the first match (operation M in CIGAR string) in the read sequence. + # The calculation is done by adding up the lengths of all the operations until the first match. + # CIGAR string is a list of tuples of (operation, length). Match is stored as CMATCH. + match_sum = 0 + for op, count in cigartuples: + if op == pysam.CMATCH: + break + match_sum += count + + first_mapped.append(match_sum) + # find which read has a cigar string that maps first than any of the + # others. + idx_min = first_mapped.index(min(first_mapped)) + + return read_list[idx_min] + + +def enlarge_bins(bin_intervals, chrom_sizes): + r""" + takes a list of consecutive but not + one after the other bin intervals + and joins them such that the + end and start of consecutive bins + is the same. + + >>> chrom_sizes = [('chr1', 100), ('chr2', 100)] + >>> bin_intervals = [('chr1', 10, 30), ('chr1', 50, 80), + ... ('chr2', 10, 60), ('chr2', 60, 90)] + >>> enlarge_bins(bin_intervals, chrom_sizes) + [('chr1', 0, 40), ('chr1', 40, 100), ('chr2', 0, 60), ('chr2', 60, 100)] + """ + # enlarge remaining bins + chr_start = True + chrom_sizes_dict = dict(chrom_sizes) + for idx in range(len(bin_intervals) - 1): + chrom, start, end = bin_intervals[idx] + chrom_next, start_next, end_next = bin_intervals[idx + 1] + if chr_start is True: + start = 0 + chr_start = False + if chrom == chrom_next and \ + end != start_next: + middle = start_next - int((start_next - end) / 2) + bin_intervals[idx] = (chrom, start, middle) + bin_intervals[idx + 1] = (chrom, middle, end_next) + if chrom != chrom_next: + bin_intervals[idx] = (chrom, start, chrom_sizes_dict[chrom]) + bin_intervals[idx + 1] = (chrom_next, 0, end_next) + + chrom, start, end = bin_intervals[-1] + bin_intervals[-1] = (chrom, start, chrom_sizes_dict[chrom]) + + return bin_intervals + + +def readBamFiles(pFileOneIterator, pFileTwoIterator, pNumberOfItemsPerBuffer, pSkipDuplicationCheck, pReadPosMatrix, pRefId2name, pMinMappingQuality): + """Read the two bam input files into n buffers each with pNumberOfItemsPerBuffer + with n = number of processes. The duplication check is handled here too.""" + buffer_mate1 = [] + buffer_mate2 = [] + duplicated_pairs = 0 + one_mate_unmapped = 0 + one_mate_not_unique = 0 + one_mate_low_quality = 0 + + all_data_read = False + j = 0 + iter_num = 0 + while j < pNumberOfItemsPerBuffer: + try: + mate1 = next(pFileOneIterator) + mate2 = next(pFileTwoIterator) + except StopIteration: + all_data_read = True + break + iter_num += 1 + + # skip 'not primary' alignments + while mate1.flag & 256 == 256: + try: + mate1 = next(pFileOneIterator) + except StopIteration: + all_data_read = True + break + while mate2.flag & 256 == 256: + try: + mate2 = next(pFileTwoIterator) + except StopIteration: + all_data_read = True + break + + assert mate1.qname == mate2.qname, "FATAL ERROR {} {} " \ + "Be sure that the sam files have the same read order " \ + "If using Bowtie2 or Hisat2 add " \ + "the --reorder option".format(mate1.qname, mate2.qname) + + # check for supplementary alignments + # (needs to be done before skipping any unmapped reads + # to keep the order of the two bam files in sync) + mate1_supplementary_list = get_supplementary_alignment( + mate1, pFileOneIterator) + mate2_supplementary_list = get_supplementary_alignment( + mate2, pFileTwoIterator) + + if mate1_supplementary_list: + mate1 = get_correct_map(mate1, mate1_supplementary_list) + + if mate2_supplementary_list: + mate2 = get_correct_map(mate2, mate2_supplementary_list) + + # skip if any of the reads is not mapped + if mate1.flag & 0x4 == 4 or mate2.flag & 0x4 == 4: + one_mate_unmapped += 1 + continue + + # skip if the read quality is low + if mate1.mapq < pMinMappingQuality or mate2.mapq < pMinMappingQuality: + # for bwa other way to test + # for multi-mapping reads is with a mapq = 0 + # the XS flag is not reliable. + if mate1.mapq == 0 & mate2.mapq == 0: + one_mate_not_unique += 1 + continue + + """ + # check if low quality is because of + # read being repetitive + # by reading the XS flag. + # The XS:i field is set by bowtie when a read is + # multi read and it contains the mapping score of the next + # best match + if 'XS' in dict(mate1.tags) or 'XS' in dict(mate2.tags): + one_mate_not_unique += 1 + continue + """ + + one_mate_low_quality += 1 + continue + + if pSkipDuplicationCheck is False: + if pReadPosMatrix.is_duplicated(pRefId2name[mate1.rname], + mate1.pos, + pRefId2name[mate2.rname], + mate2.pos): + duplicated_pairs += 1 + continue + buffer_mate1.append(mate1) + buffer_mate2.append(mate2) + j += 1 + + if all_data_read and len(buffer_mate1) != 0 and len(buffer_mate2) != 0: + return buffer_mate1, buffer_mate2, True, duplicated_pairs, one_mate_unmapped, one_mate_not_unique, one_mate_low_quality, iter_num - len(buffer_mate1) + if all_data_read and len(buffer_mate1) == 0 or len(buffer_mate2) == 0: + return None, None, True, duplicated_pairs, one_mate_unmapped, one_mate_not_unique, one_mate_low_quality, iter_num - len(buffer_mate1) + return buffer_mate1, buffer_mate2, False, duplicated_pairs, one_mate_unmapped, one_mate_not_unique, one_mate_low_quality, iter_num - len(buffer_mate1) + + + +def process_data(pMateBuffer1, pMateBuffer2, pMinMappingQuality, + pKeepSelfCircles, pRestrictionSequence, pKeepSelfLigation, pMatrixSize, + pRfPositions, pRefId2name, + pDanglingSequences, pBinsize, pResultIndex, + pQueueOut, pTemplate, pOutputBamSet, pCounter, + pSharedBinIntvalTree, pDictBinIntervalTreeIndex, pCoverage, pCoverageIndex, + pOutputFileBufferDir, pRow, pCol, pData, + pMaxInsertSize, pQuickQCMode): + """ + This function computes for a given number of elements in pMateBuffer1 and pMaterBuffer2 a partial interaction matrix. + This function is used by multiple processes to speed up the computation. + All partial matrices are merged in the end into one interaction matrix. + + Parameters + ---------- + pMateBuffer1 : List of n reads of type 'pysam.libcalignedsegment.AlignedSegment' of sam input file 1 + pMateBuffer2 : List of n reads of type 'pysam.libcalignedsegment.AlignedSegment' of sam input file 2 + pMinMappingQuality : integer, minimum mapping quality of a read + pKeepSelfCircles : boolean, if self circles should be kept + pRestrictionSequence : List of String, the restriction sequence + pKeepSelfLigation : If self ligations should be removed + pMatrixSize : integer, the size of the interaction matrix + pRfPositions : intervalTree, only used if a restriction cut file and not a bin size was defined. + pRefId2name : Tuple, Maps a reference id to a name + pDanglingSequences : dict, dict of dangling sequences + pBinsize : integer, the size of the bins + pResultIndex : integer, number of processs, range(0, threads). Is returned via the queue to have access to the right row, col and data array after the computation. + pQueueOut : multiprocessing.Queue, queue to return the computed counting variables: + one_mate_unmapped, one_mate_low_quality, one_mate_not_unique, dangling_end, self_circle, self_ligation, same_fragment, + mate_not_close_to_rf, count_inward, count_outward, count_left, count_right, inter_chromosomal, short_range, long_range, + pair_added, len(pMateBuffer1), pResultIndex, pCounter + pTemplate : The template for the output bam file + pOutputBamSet : If a output bam file should be written. Depending on the input parameter '--outBam' + pOutputName : String, Name of the partial bam file + pCounter : integer, value which is returned to the main process. The main process can than write a pCounter.bam_done file + to signal the background process, which is merging the partial bam files into one, that this dataset can be merged. + pSharedBinIntvalTree : multiprocessing.sharedctype.RawArray of C_Interval, stores the interval tree in a 1D-RawArray. + pDictBinIntervalTreeIndex : dict, stores the information at which index position a given interval starts and ends in the 1D-array 'pSharedBinIntvalTree' + pCoverage : multiprocessing.sharedctype.Array of c_uint, Stores the coverage in a 1D-Array + pCoverageIndex : multiprocessing.sharedctype.RawArray of C_Coverage, stores the information in the 1D-array 'pCoverage' + pOutputFileBufferDir : String, the directory where the partial output bam files are buffered. Default is '/dev/shm/' + pRow : multiprocessing.sharedctype.RawArray of c_uint, Stores the row index information. It is available for all processes and does not need to be copied. + pCol : multiprocessing.sharedctype.RawArray of c_uint, stores the column index information. It is available for all processes and does not need to be copied. + pData : multiprocessing.sharedctype.RawArray of c_ushort, stores a 1 for each row - column pair. It is available for all processes and does not need to be copied. + pMaxInsertSize : maximum illumina insert size + """ + try: + + one_mate_unmapped = 0 + one_mate_low_quality = 0 + one_mate_not_unique = 0 + dangling_end = {} + if pRestrictionSequence is not None: + for restrictionSequence in pRestrictionSequence: + dangling_end[restrictionSequence] = 0 + self_circle = 0 + self_ligation = 0 + same_fragment = 0 + mate_not_close_to_rf = 0 + + count_inward = 0 + count_outward = 0 + count_left = 0 + count_right = 0 + inter_chromosomal = 0 + short_range = 0 + long_range = 0 + + pair_added = 0 + + iter_num = 0 + hic_matrix = None + + out_bam_index_buffer = [] + + if pMateBuffer1 is None or pMateBuffer2 is None: + + pQueueOut.put([hic_matrix, [one_mate_unmapped, one_mate_low_quality, one_mate_not_unique, dangling_end, self_circle, self_ligation, same_fragment, + mate_not_close_to_rf, count_inward, count_outward, + count_left, count_right, inter_chromosomal, short_range, long_range, pair_added, iter_num, pResultIndex, out_bam_index_buffer]]) + return + + while iter_num < len(pMateBuffer1) and iter_num < len(pMateBuffer2): + mate1 = pMateBuffer1[iter_num] + mate2 = pMateBuffer2[iter_num] + iter_num += 1 + + # check if reads belong to a bin + # + # pDictBinInterval stores the start and end position for each chromsome in the array 'pSharedBinIntvalTree' + # To get to the right interval a binary search is used. + mate_bins = [] + mate_is_unasigned = False + for mate in [mate1, mate2]: + mate_ref = pRefId2name[mate.rname] + # find the middle genomic position of the read. This is used to + # find the bin it belongs to. + read_middle = mate.pos + int(mate.qlen / 2) + try: + start, end = pDictBinIntervalTreeIndex[mate_ref] + middle_pos = int((start + end) / 2) + mate_bin = None + while not start > end: + if pSharedBinIntvalTree[middle_pos].begin <= read_middle and read_middle <= pSharedBinIntvalTree[middle_pos].end: + mate_bin = pSharedBinIntvalTree[middle_pos] + mate_is_unasigned = False + break + elif pSharedBinIntvalTree[middle_pos].begin > read_middle: + end = middle_pos - 1 + middle_pos = int((start + end) / 2) + mate_is_unasigned = True + else: + start = middle_pos + 1 + middle_pos = int((start + end) / 2) + mate_is_unasigned = True + + except KeyError: + # for small contigs it can happen that they are not + # in the bin_intval_tree keys if no restriction site is found + # on the contig. + mate_is_unasigned = True + break + + # report no match case + if mate_bin is None: + mate_is_unasigned = True + break + mate_bin_id = mate_bin.data + mate_bins.append(mate_bin_id) + + # if a mate is unassigned, it means it is not close + # to a restriction site + if mate_is_unasigned is True: + mate_not_close_to_rf += 1 + continue + + # check if mates are in the same chromosome + if mate1.reference_id != mate2.reference_id: + orientation = 'diff_chromosome' + else: + # to identify 'inward' and 'outward' orientations + # the order or the mates in the genome has to be + # known. + if mate1.pos < mate2.pos: + first_mate = mate1 + second_mate = mate2 + else: + first_mate = mate2 + second_mate = mate1 + + """ + outward + <--------------- ----------------> + + inward + ---------------> <---------------- + + same-strand-right + ---------------> ----------------> + + same-strand-left + <--------------- <---------------- + """ + + if not first_mate.is_reverse and second_mate.is_reverse: + orientation = 'inward' + elif first_mate.is_reverse and not second_mate.is_reverse: + orientation = 'outward' + elif first_mate.is_reverse and second_mate.is_reverse: + orientation = 'same-strand-left' + else: + orientation = 'same-strand-right' + + # check self-circles + # self circles are defined as outward pairs that do not + # have a restriction sequence in between. The distance of < 25kb is + # used to only check close outward pairs as far apart pairs can not be self-circles + if abs(mate2.pos - mate1.pos) < 25000 and orientation == 'outward': + has_rf = [] + + if pRfPositions and pRestrictionSequence: + # check if in between the two mate + # ends the restriction fragment is found. + # check for multiple restriction sequences + for restrictionSequence in pRestrictionSequence: + # the interval used is: + # start of fragment + length of restriction sequence + # end of fragment - length of restriction sequence + # the restriction sequence length is subtracted + # such that only fragments internally containing + # the restriction site are identified + frag_start = min(mate1.pos, mate2.pos) + \ + len(restrictionSequence) + frag_end = max(mate1.pos + mate1.qlen, mate2.pos + mate2.qlen) - len(restrictionSequence) + mate_ref = pRefId2name[mate1.rname] + if mate_ref in pRfPositions: + has_rf.extend(sorted( + pRfPositions[mate_ref][frag_start: frag_end])) + + if len(has_rf) == 0: + self_circle += 1 + if not pKeepSelfCircles: + continue + if abs(mate2.pos - mate1.pos) < pMaxInsertSize and orientation == 'inward': + # check for dangling ends if the restriction sequence is known and if they look + # like 'same fragment' + if pRestrictionSequence: + if pDanglingSequences: + + # check for dangling ends in sequence. Stop check with first match + one_match = False + for restrictionSequence in pRestrictionSequence: + if check_dangling_end(mate1, pDanglingSequences[restrictionSequence]) or \ + check_dangling_end(mate2, pDanglingSequences[restrictionSequence]): + dangling_end[restrictionSequence] += 1 + one_match = True + break + if one_match: + continue + has_rf = [] + + if pRfPositions and pRestrictionSequence: + # check if in between the two mate + # ends the restriction fragment is found. + + # the interval used is: + # start of fragment + length of restriction sequence + # end of fragment - length of restriction sequence + # the restriction sequence length is subtracted + # such that only fragments internally containing + # the restriction site are identified + + for restrictionSequence in pRestrictionSequence: + frag_start = min(mate1.pos, mate2.pos) + \ + len(restrictionSequence) + frag_end = max(mate1.pos + mate1.qlen, mate2.pos + mate2.qlen) - len(restrictionSequence) + mate_ref = pRefId2name[mate1.rname] + if mate_ref in pRfPositions: + has_rf.extend(sorted( + pRfPositions[mate_ref][frag_start: frag_end])) + + # case when there is no restriction fragment site between the + # mates + if len(has_rf) == 0: + same_fragment += 1 + continue + + self_ligation += 1 + + if not pKeepSelfLigation: + # skip self ligations + continue + + # set insert size to save bam + mate1.isize = mate2.pos - mate1.pos + mate2.isize = mate1.pos - mate2.pos + + # if mate_bins, which is set in the previous section + # does not have size=2, it means that one + # of the exceptions happened + if len(mate_bins) != 2: + continue + + # count type of pair (distance, orientation) + if mate1.reference_id != mate2.reference_id: + inter_chromosomal += 1 + + elif abs(mate2.pos - mate1.pos) < 20000: + short_range += 1 + else: + long_range += 1 + + if orientation == 'inward': + count_inward += 1 + elif orientation == 'outward': + count_outward += 1 + elif orientation == 'same-strand-left': + count_left += 1 + elif orientation == 'same-strand-right': + count_right += 1 + + for mate in [mate1, mate2]: + # fill in coverage vector + vec_start = int(max(0, mate.pos - mate_bin.begin) / pBinsize) + length_coverage = pCoverageIndex[mate_bin_id].end - \ + pCoverageIndex[mate_bin_id].begin + vec_end = min(length_coverage, int( + vec_start + len(mate.seq) / pBinsize)) + coverage_index = pCoverageIndex[mate_bin_id].begin + vec_start + coverage_end = pCoverageIndex[mate_bin_id].begin + vec_end + for i in range(coverage_index, coverage_end, 1): + pCoverage[i] += 1 + + if not pQuickQCMode: + pRow[pair_added] = mate_bins[0] + pCol[pair_added] = mate_bins[1] + pData[pair_added] = np.uint8(1) + + pair_added += 1 + if pOutputBamSet: + + out_bam_index_buffer.append(iter_num - 1) + + pQueueOut.put([[one_mate_unmapped, one_mate_low_quality, one_mate_not_unique, dangling_end, self_circle, self_ligation, same_fragment, + mate_not_close_to_rf, count_inward, count_outward, + count_left, count_right, inter_chromosomal, short_range, long_range, pair_added, len(pMateBuffer1), pResultIndex, pCounter, out_bam_index_buffer]]) + except Exception as exp: + pQueueOut.put('Fail: ' + str(exp) + traceback.format_exc()) + return + return + +def createMatrix(pOutFileName, pMaxDistance, pMaxLibraryInsertSize, pQCfolder, + pThreads, pDanglingSequence, pRestrictionSequence, pSamFiles, + pDoTestRun, pOutBam, pChromosomeSizes, pRestrictionCutFile, + pRegion, pBinSize, pInputBufferSize, + pDoTestRunLines, pSkipDuplicationCheck, pMinMappingQuality, + pKeepSelfCircles, pKeepSelfLigation, pMinDistance, pGenomeAssembly): + # pOutFileName.name = pOutFileName.name.strip() + # log.debug('pOutFileName.name: {}'.format(pOutFileName.name.endswith('.h5'))) + if not pOutFileName.name.endswith('.h5') and not pOutFileName.name.endswith('.cool'): + if '.mcool' not in pOutFileName.name: + log.error('Please define the file extension. h5 and cool are supported, or the specializations of cool, mcool. Given input {}'.format(pOutFileName.name)) + exit(1) + # for backwards compatibility + if pMaxDistance is not None: + pMaxLibraryInsertSize = pMaxDistance + try: + QC.make_sure_path_exists(pQCfolder) + except OSError: + exit("Can't open/create QC folder path: {}. Please check".format(pQCfolder)) + + if pThreads < 2: + pThreads = 2 + warnings.warn( + "\nAt least two threads need to be defined. Setting --threads = 2!s\n") + + if pDanglingSequence and not pRestrictionSequence: + exit("\nIf --danglingSequence is set, --restrictionSequence needs to be set too.\n") + + log.info("reading {} and {} to build hic_matrix\n".format(pSamFiles[0].name, + pSamFiles[1].name)) + str1 = pysam.Samfile(pSamFiles[0].name, 'rb') + str2 = pysam.Samfile(pSamFiles[1].name, 'rb') + + pSamFiles[0].close() + pSamFiles[1].close() + if not pDoTestRun: + if pOutBam: + pOutBam.close() + out_bam_file = pysam.Samfile(pOutBam.name, 'wb', template=str1) + + if pChromosomeSizes is None: + chrom_sizes = get_chrom_sizes(str1) + else: + chrom_sizes = OrderedDict() + with open(pChromosomeSizes.name, 'r') as file: + file_ = True + while file_: + file_ = file.readline().strip() + if file_ != '': + line_split = file_.split('\t') + chrom_sizes[line_split[0]] = int(line_split[1]) + chrom_sizes = list(chrom_sizes.items()) + + # log.debug('chrom_sizes {}'.format(chrom_sizes)) + read_pos_matrix = ReadPositionMatrix() + + rf_interval = [] + if pRestrictionCutFile: + for restrictionCutFile in pRestrictionCutFile: + rf_interval.extend(bed2interval_list(restrictionCutFile, chrom_sizes, pRegion)) + + rf_positions = intervalListToIntervalTree(rf_interval) + log.debug('rf_positions {}'.format(rf_positions.keys())) + else: + rf_positions = None + + if pBinSize: + bin_intervals = get_bins(pBinSize[0], chrom_sizes, pRegion) + else: + bin_intervals = get_rf_bins(rf_interval, + min_distance=pMinDistance, + max_distance=pMaxLibraryInsertSize) + + matrix_size = len(bin_intervals) + bin_intval_tree = intervalListToIntervalTree(bin_intervals) + ref_id2name = str1.references + + # build c_type shared memory for the interval tree + shared_array_list = [] + index_dict = {} + end = -1 + for i, seq in enumerate(bin_intval_tree): + start = end + 1 + interval_list = [] + for interval in bin_intval_tree[seq]: + interval_list.append((interval.begin, interval.end, interval.data)) + end = start + len(bin_intval_tree[seq]) - 1 + index_dict[seq] = (start, end) + interval_list = sorted(interval_list) + shared_array_list.extend(interval_list) + shared_build_intval_tree = RawArray(C_Interval, shared_array_list) + bin_intval_tree = None + dangling_sequences = {} + if pDanglingSequence: + # build a list of dangling sequences + if pRestrictionSequence is not None: + for i in range(0, len(pRestrictionSequence)): + pRestrictionSequence[i] = pRestrictionSequence[i].upper() + pDanglingSequence[i] = pDanglingSequence[i].upper() + dangling_sequences[pRestrictionSequence[i]] = {} + dangling_sequences[pRestrictionSequence[i]]['pat_forw'] = pDanglingSequence[i] + dangling_sequences[pRestrictionSequence[i]]['pat_rev'] = str( + Seq(pDanglingSequence[i]).reverse_complement()) + + log.info("dangling sequences to check " + "are {}\n".format(dangling_sequences)) + + # initialize coverage vectors that + # save the number of reads that overlap + # a bin. + # To save memory, coverage is not measured by bp + # but by bins of length 10bp + binsize = 10 + number_of_elements_coverage = 0 + start_pos_coverage = [] + end_pos_coverage = [] + + for chrom, start, end in bin_intervals: + start_pos_coverage.append(number_of_elements_coverage) + + number_of_elements_coverage += (end - start) // binsize + end_pos_coverage.append(number_of_elements_coverage - 1) + pos_coverage = RawArray(C_Coverage, list(zip( + start_pos_coverage, end_pos_coverage))) + start_pos_coverage = None + end_pos_coverage = None + coverage = Array(c_uint, [0] * number_of_elements_coverage) + + # define global shared ctypes arrays for row, col and data + pThreads = pThreads - 1 + row = [None] * pThreads + col = [None] * pThreads + data = [None] * pThreads + for i in range(pThreads): + row[i] = RawArray(c_uint, pInputBufferSize) + col[i] = RawArray(c_uint, pInputBufferSize) + data[i] = RawArray(c_ushort, pInputBufferSize) + + start_time = time.time() + + iter_num = 0 + # pair_added = 0 + # hic_matrix = None + + one_mate_unmapped = 0 + one_mate_low_quality = 0 + one_mate_not_unique = 0 + dangling_end = {} + if pRestrictionSequence is not None: + for restrictionSequence in pRestrictionSequence: + dangling_end[restrictionSequence] = 0 + + self_circle = 0 + self_ligation = 0 + same_fragment = 0 + mate_not_close_to_rf = 0 + duplicated_pairs = 0 + + count_inward = 0 + count_outward = 0 + count_left = 0 + count_right = 0 + inter_chromosomal = 0 + short_range = 0 + long_range = 0 + + pair_added = 0 + + # input buffer for bam files + buffer_workers1 = [None] * pThreads + buffer_workers2 = [None] * pThreads + + # output buffer to write bam with mate1 and mate2 pairs + process = [None] * pThreads + all_data_processed = False + hic_matrix = coo_matrix((matrix_size, matrix_size), dtype='uint32') + queue = [None] * pThreads + + all_threads_done = False + thread_done = [False] * pThreads + count_output = 0 + count_call_of_read_input = 0 + computed_pairs = 0 + + if pDoTestRun: + pInputBufferSize = pDoTestRunLines + fail_flag = False + fail_message = '' + while not all_data_processed or not all_threads_done: + + for i in range(pThreads): + if queue[i] is None and not all_data_processed: + count_call_of_read_input += 1 + + buffer_workers1[i], buffer_workers2[i], all_data_processed, \ + duplicated_pairs_, one_mate_unmapped_, one_mate_not_unique_, \ + one_mate_low_quality_, iter_num_ = readBamFiles(pFileOneIterator=str1, + pFileTwoIterator=str2, + pNumberOfItemsPerBuffer=pInputBufferSize, + pSkipDuplicationCheck=pSkipDuplicationCheck, + pReadPosMatrix=read_pos_matrix, + pRefId2name=ref_id2name, + pMinMappingQuality=pMinMappingQuality + ) + duplicated_pairs += duplicated_pairs_ + one_mate_unmapped += one_mate_unmapped_ + one_mate_not_unique += one_mate_not_unique_ + one_mate_low_quality += one_mate_low_quality_ + iter_num += iter_num_ + queue[i] = Queue() + thread_done[i] = False + computed_pairs += len(buffer_workers1[i]) + # create process to compute hic matrix for this buffer + process[i] = Process(target=process_data, kwargs=dict( + pMateBuffer1=buffer_workers1[i], + pMateBuffer2=buffer_workers2[i], + pMinMappingQuality=pMinMappingQuality, + pKeepSelfCircles=pKeepSelfCircles, + pRestrictionSequence=pRestrictionSequence, + pKeepSelfLigation=pKeepSelfLigation, + pMatrixSize=matrix_size, + pRfPositions=rf_positions, + pRefId2name=ref_id2name, + pDanglingSequences=dangling_sequences, + pBinsize=binsize, + pResultIndex=i, + pQueueOut=queue[i], + pTemplate=str1, + pOutputBamSet=pOutBam, + pCounter=count_output, + pSharedBinIntvalTree=shared_build_intval_tree, + pDictBinIntervalTreeIndex=index_dict, + pCoverage=coverage, + pCoverageIndex=pos_coverage, + pOutputFileBufferDir="", + pRow=row[i], + pCol=col[i], + pData=data[i], + pMaxInsertSize=pMaxLibraryInsertSize, + pQuickQCMode=pDoTestRun + )) + process[i].start() + count_output += 1 + + elif queue[i] is not None and not queue[i].empty(): + result = queue[i].get() + if 'Fail:' in result: + fail_flag = True + fail_message = result[6:] + else: + if result[0] is not None: + elements = result[0][15] + if hic_matrix is None: + hic_matrix = coo_matrix( + (data[i][:elements], (row[i][:elements], col[i][:elements])), shape=(matrix_size, matrix_size)) + else: + hic_matrix += coo_matrix( + (data[i][:elements], (row[i][:elements], col[i][:elements])), shape=(matrix_size, matrix_size)) + + for sequence in result[0][3]: + dangling_end[sequence] += result[0][3][sequence] + self_circle += result[0][4] + self_ligation += result[0][5] + same_fragment += result[0][6] + mate_not_close_to_rf += result[0][7] + + count_inward += result[0][8] + count_outward += result[0][9] + count_left += result[0][10] + count_right += result[0][11] + inter_chromosomal += result[0][12] + short_range += result[0][13] + long_range += result[0][14] + + pair_added += result[0][15] + iter_num += result[0][16] + + for bam_index in result[0][19]: + mate1 = buffer_workers1[i][bam_index] + mate2 = buffer_workers2[i][bam_index] + + mate1.flag |= 0x1 + mate2.flag |= 0x1 + + # set one read as the first in pair and the + # other as second + mate1.flag |= 0x40 + mate2.flag |= 0x80 + + # set chrom of mate + mate1.mrnm = mate2.rname + mate2.mrnm = mate1.rname + + # set position of mate + mate1.mpos = mate2.pos + mate2.mpos = mate1.pos + + out_bam_file.write(mate1) + out_bam_file.write(mate2) + + buffer_workers1[i] = None + buffer_workers2[i] = None + queue[i] = None + process[i].join() + process[i].terminate() + process[i] = None + thread_done[i] = True + + # caused by the architecture I try to display this output + # information after +-1e5 of 1e6 reads. + if iter_num % 1e6 < 100000: + elapsed_time = time.time() - start_time + log.info("processing {} lines took {:.2f} " + "secs ({:.1f} lines per " + "second)\n".format(iter_num, + elapsed_time, + iter_num / elapsed_time)) + log.info("{} ({:.2f}%) valid pairs added to matrix" + "\n".format(pair_added, float(100 * pair_added) / iter_num)) + if pDoTestRun and iter_num > pDoTestRunLines: + log.debug( + "\n## *WARNING*. Early exit because of --doTestRun parameter ##\n\n") + all_data_processed = True + thread_done[i] = True + break + elif all_data_processed and queue[i] is None: + thread_done[i] = True + else: + time.sleep(1) + + if all_data_processed: + all_threads_done = True + for thread in thread_done: + if not thread: + all_threads_done = False + if fail_flag: + log.error(fail_message) + exit(1) + else: + log.debug('Parallel stuff done') + if not pDoTestRun: + # the resulting matrix is only filled unevenly with some pairs + # int the upper triangle and others in the lower triangle. To construct + # the definite matrix I add the values from the upper and lower triangles + # and subtract the diagonal to avoid double counting it. + # The resulting matrix is symmetric. + if pOutBam: + out_bam_file.close() + + dia = dia_matrix(([hic_matrix.diagonal()], [0]), + shape=hic_matrix.shape) + hic_matrix = hic_matrix + hic_matrix.T - dia + # extend bins such that they are next to each other + bin_intervals = enlarge_bins(bin_intervals[:], chrom_sizes) + # compute max bin coverage + bin_max = [] + + for cover in pos_coverage: + max_element = 0 + for i in range(cover.begin, cover.end, 1): + if coverage[i] > max_element: + max_element = coverage[i] + if max_element == 0: + bin_max.append(np.nan) + else: + bin_max.append(max_element) + + chr_name_list, start_list, end_list = list(zip(*bin_intervals)) + bin_intervals = list(zip(chr_name_list, start_list, end_list, bin_max)) + hic_ma = hm.hiCMatrix() + hic_ma.setMatrix(hic_matrix, cut_intervals=bin_intervals) + + """ + if pRestrictionCutFile: + # load the matrix to mask those + # bins that most likely didn't + # have a restriction site that was cutted + + # reload the matrix as a Hi-C matrix object + hic_matrix = hm.hiCMatrix(pOutFileName.name) + + hic_matrix.maskBins(get_poor_bins(bin_max)) + hic_matrix.save(pOutFileName.name) + """ + if not pKeepSelfLigation: + msg = " (removed)" + else: + msg = " (not removed)" + + mappable_unique_high_quality_pairs = iter_num - \ + (one_mate_unmapped + one_mate_low_quality + one_mate_not_unique) + + intermediate_qc_log = StringIO() + + if pMinDistance: + intermediate_qc_log.write(""" + File\t{}\t\t + Sequenced reads\t{}\t\t + Min rest. site distance\t{}\t\t + Max library insert size\t{}\t\t + + """.format(pOutFileName.name, iter_num, pMinDistance, pMaxLibraryInsertSize)) + else: + intermediate_qc_log.write(""" + File\t{}\t\t + Sequenced reads\t{}\t\t + Max library insert size\t{}\t\t + + """.format(pOutFileName.name, iter_num, pMaxLibraryInsertSize)) + + intermediate_qc_log.write( + "#\tcount\t(percentage w.r.t. total sequenced reads)\n") + + intermediate_qc_log.write("Pairs mappable, unique and high quality\t{}\t({:.2f})\n". + format(mappable_unique_high_quality_pairs, + 100 * float(mappable_unique_high_quality_pairs) / iter_num)) + + intermediate_qc_log.write("Hi-C contacts\t{}\t({:.2f})\n". + format(pair_added, 100 * float(pair_added) / iter_num)) + + intermediate_qc_log.write("One mate unmapped\t{}\t({:.2f})\n". + format(one_mate_unmapped, 100 * float(one_mate_unmapped) / iter_num)) + + intermediate_qc_log.write("One mate not unique\t{}\t({:.2f})\n". + format(one_mate_not_unique, 100 * float(one_mate_not_unique) / iter_num)) + + intermediate_qc_log.write("Low mapping quality\t{}\t({:.2f})\n". + format(one_mate_low_quality, 100 * float(one_mate_low_quality) / iter_num)) + + intermediate_qc_log.write( + "\n#\tcount\t(percentage w.r.t. mappable, unique and high quality pairs)\n") + + if len(dangling_end) > 0: + log.debug('dangling_sequences {}'.format(dangling_sequences.keys())) + log.debug('dangling_end {}'.format(dangling_end.keys())) + # log.debug('dangling_end {}'.format(res.keys())) + + for key in dangling_end: + # dangling_sequences[pRestrictionSequence[i]]['pat_forw'] + intermediate_qc_log.write("dangling end {} (restriction sequence {})\t{}\t({:.2f})\n". + format(dangling_sequences[key]['pat_forw'], key, dangling_end[key], 100 * float(dangling_end[key]) / mappable_unique_high_quality_pairs)) + else: + intermediate_qc_log.write("dangling end\t{}\t({:.2f})\n". + format(0, 100 * float(0) / mappable_unique_high_quality_pairs)) + if pRestrictionCutFile is not None: + intermediate_qc_log.write("self ligation{}\t{}\t({:.2f})\n". + format(msg, self_ligation, 100 * float(self_ligation) / mappable_unique_high_quality_pairs)) + + intermediate_qc_log.write("One mate not close to rest site\t{}\t({:.2f})\n". + format(mate_not_close_to_rf, 100 * float(mate_not_close_to_rf) / mappable_unique_high_quality_pairs)) + intermediate_qc_log.write("self circle\t{}\t({:.2f})\n". + format(self_circle, 100 * float(self_circle) / mappable_unique_high_quality_pairs)) + + intermediate_qc_log.write("same fragment\t{}\t({:.2f})\n". + format(same_fragment, 100 * float(same_fragment) / mappable_unique_high_quality_pairs)) + + intermediate_qc_log.write("duplicated pairs\t{}\t({:.2f})\n". + format(duplicated_pairs, 100 * float(duplicated_pairs) / mappable_unique_high_quality_pairs)) + + if pair_added > 0: + intermediate_qc_log.write( + "\n#\tcount\t(percentage w.r.t. total valid pairs used)\n") + intermediate_qc_log.write("inter chromosomal\t{}\t({:.2f})\n". + format(inter_chromosomal, 100 * float(inter_chromosomal) / pair_added)) + + intermediate_qc_log.write("Intra short range (< 20kb)\t{}\t({:.2f})\n". + format(short_range, 100 * float(short_range) / pair_added)) + + intermediate_qc_log.write("Intra long range (>= 20kb)\t{}\t({:.2f})\n". + format(long_range, 100 * float(long_range) / pair_added)) + + intermediate_qc_log.write("Read pair type: inward pairs\t{}\t({:.2f})\n". + format(count_inward, 100 * float(count_inward) / pair_added)) + + intermediate_qc_log.write("Read pair type: outward pairs\t{}\t({:.2f})\n". + format(count_outward, 100 * float(count_outward) / pair_added)) + + intermediate_qc_log.write("Read pair type: left pairs\t{}\t({:.2f})\n". + format(count_left, 100 * float(count_left) / pair_added)) + + intermediate_qc_log.write("Read pair type: right pairs\t{}\t({:.2f})\n". + format(count_right, 100 * float(count_right) / pair_added)) + + log_file_name = os.path.join(pQCfolder, "QC.log") + log_file = open(log_file_name, 'w') + log_file.write(intermediate_qc_log.getvalue()) + log_file.close() + log.debug('log_file_name {}'.format(log_file_name)) + log.debug('pQCfolder {}'.format(pQCfolder)) + + QC.main("-l {} -o {}".format(log_file_name, pQCfolder).split()) + + pOutFileName.close() + # removing the empty file. Otherwise the save method + # will say that the file already exists. + unlink(pOutFileName.name) + + hic_metadata = {} + hic_metadata['statistics'] = intermediate_qc_log.getvalue() + hic_metadata['matrix-generated-by'] = np.string_( + 'HiCExplorer-' + __version__) + hic_metadata['matrix-generated-by-url'] = np.string_( + 'https://github.com/deeptools/HiCExplorer') + if pGenomeAssembly: + hic_metadata['genome-assembly'] = np.string_(pGenomeAssembly) + + intermediate_qc_log.close() + if pOutFileName.name.endswith('.mcool') and pBinSize is not None and len(pBinSize) > 2: + + matrixFileHandlerOutput = MatrixFileHandler( + pFileType='cool', pHiCInfo=hic_metadata) + matrixFileHandlerOutput.set_matrix_variables(hic_ma.matrix, + hic_ma.cut_intervals, + hic_ma.nan_bins, + hic_ma.correction_factors, + hic_ma.distance_counts) + matrixFileHandlerOutput.save(pOutFileName.name + '::/resolutions/' + str( + pBinSize[0]), pSymmetric=True, pApplyCorrection=False) + + for resolution in pBinSize[1:]: + hic_matrix_to_merge = deepcopy(hic_ma) + _mergeFactor = int(resolution) // pBinSize[0] + merged_matrix = hicMergeMatrixBins.merge_bins( + hic_matrix_to_merge, _mergeFactor) + matrixFileHandlerOutput = MatrixFileHandler( + pFileType='cool', pAppend=True, pHiCInfo=hic_metadata) + matrixFileHandlerOutput.set_matrix_variables(merged_matrix.matrix, + merged_matrix.cut_intervals, + merged_matrix.nan_bins, + merged_matrix.correction_factors, + merged_matrix.distance_counts) + matrixFileHandlerOutput.save(pOutFileName.name + '::/resolutions/' + str( + resolution), pSymmetric=True, pApplyCorrection=False) + + else: + if not pDoTestRun: + hic_ma.save(pOutFileName.name, pHiCInfo=hic_metadata) diff --git a/hicexplorer/test/general/test_hicBuildMatrixMicroC.py b/hicexplorer/test/general/test_hicBuildMatrixMicroC.py new file mode 100644 index 00000000..c1fdbd0f --- /dev/null +++ b/hicexplorer/test/general/test_hicBuildMatrixMicroC.py @@ -0,0 +1,321 @@ +import warnings +warnings.simplefilter(action="ignore", category=RuntimeWarning) +warnings.simplefilter(action="ignore", category=PendingDeprecationWarning) +from hicexplorer import hicBuildMatrixMicroC, hicInfo +from hicmatrix import HiCMatrix as hm +from tempfile import NamedTemporaryFile, mkdtemp +import shutil +import os +import numpy.testing as nt +import pytest +from hicexplorer.test.test_compute_function import compute + +ROOT = os.path.join(os.path.dirname(os.path.dirname(os.path.abspath(__file__))), "test_data/") +sam_R1 = ROOT + "small_test_R1_unsorted.bam" +sam_R2 = ROOT + "small_test_R2_unsorted.bam" +delta = 80000 + + +def are_files_equal(file1, file2, delta=1): + equal = True + if delta: + mismatches = 0 + with open(file1) as textfile1, open(file2) as textfile2: + for x, y in zip(textfile1, textfile2): + if x.startswith('File'): + continue + if x != y: + if delta: + mismatches += 1 + if mismatches > delta: + equal = False + break + else: + equal = False + break + return equal + + +def test_build_matrix(capsys): + outfile = NamedTemporaryFile(suffix='.h5', delete=False) + outfile.close() + outfile_bam = NamedTemporaryFile(suffix='.bam', delete=False) + outfile.close() + qc_folder = mkdtemp(prefix="testQC_") + args = "-s {} {} --outFileName {} -bs 5000 -b {} --QCfolder {} --threads 4".format(sam_R1, sam_R2, + outfile.name, outfile_bam.name, + qc_folder).split() + # hicBuildMatrixMicroC.main(args) + compute(hicBuildMatrixMicroC.main, args, 5) + test = hm.hiCMatrix(ROOT + "small_test_matrix_parallel_one_rc.h5") + new = hm.hiCMatrix(outfile.name) + nt.assert_equal(test.matrix.data, new.matrix.data) + nt.assert_equal(test.cut_intervals, new.cut_intervals) + # print("MATRIX NAME:", outfile.name) + print(set(os.listdir(ROOT + "QC/"))) + assert are_files_equal(ROOT + "QC/QC.log", qc_folder + "/QC.log") + assert set(os.listdir(ROOT + "QC/")) == set(os.listdir(qc_folder)) + + # accept delta of 80 kb, file size is around 4.5 MB + assert abs(os.path.getsize(ROOT + "small_test_matrix_result.bam") - os.path.getsize(outfile_bam.name)) < delta + + os.unlink(outfile.name) + shutil.rmtree(qc_folder) + # os.unlink("/tmp/test.bam") + + +# def test_build_matrix_restriction_enzyme_region(capsys): +# outfile = NamedTemporaryFile(suffix='.h5', delete=False) +# outfile.close() +# outfile_bam = NamedTemporaryFile(suffix='.bam', delete=False) +# outfile.close() +# qc_folder = mkdtemp(prefix="testQC_") +# args = "-s {} {} --outFileName {} -bs 5000 -b {} --QCfolder {} --threads 4 --region {}".format(sam_R1, sam_R2, outfile.name, outfile_bam.name, qc_folder, 'chr3R').split() +# # hicBuildMatrixMicroC.main(args) +# compute(hicBuildMatrixMicroC.main, args, 5) + +# test = hm.hiCMatrix(ROOT + "small_test_matrix_parallel_two_rc_chr3R.h5") +# new = hm.hiCMatrix(outfile.name) + +# # print('test.cut_intervals {}'.format(test.cut_intervals)) +# # print('new.cut_intervals {}'.format(new.cut_intervals)) +# nt.assert_equal(test.matrix.data, new.matrix.data) +# nt.assert_equal(len(test.cut_intervals), len(new.cut_intervals)) + +# # print("MATRIX NAME:", outfile.name) +# print(set(os.listdir(ROOT + "QC_region/"))) +# assert are_files_equal(ROOT + "QC_region/QC.log", qc_folder + "/QC.log") +# assert set(os.listdir(ROOT + "QC_region/")) == set(os.listdir(qc_folder)) + +# # accept delta of 80 kb, file size is around 4.5 MB +# assert abs(os.path.getsize(ROOT + "build_region.bam") - os.path.getsize(outfile_bam.name)) < delta + +# os.unlink(outfile.name) +# shutil.rmtree(qc_folder) + + +# def test_build_matrix_chromosome_sizes(capsys): +# outfile = NamedTemporaryFile(suffix='.h5', delete=False) +# outfile.close() +# outfile_bam = NamedTemporaryFile(suffix='.bam', delete=False) +# outfile.close() +# qc_folder = mkdtemp(prefix="testQC_") +# args = "-s {} {} --outFileName {} -bs 5000 -b {} --QCfolder {} --threads 4 --chromosomeSizes {} \ +# --restrictionSequence GATC --danglingSequence GATC -rs {}".format(sam_R1, sam_R2, +# outfile.name, outfile_bam.name, +# qc_folder, ROOT + 'hicBuildMatrixMicroC/dm3.chrom.sizes', dpnii_file).split() +# # hicBuildMatrixMicroC.main(args) +# compute(hicBuildMatrixMicroC.main, args, 5) + +# test = hm.hiCMatrix(ROOT + "hicBuildMatrixMicroC/chromosome_sizes/small_test_chromosome_size.h5") +# new = hm.hiCMatrix(outfile.name) +# nt.assert_equal(test.matrix.data, new.matrix.data) +# nt.assert_equal(test.cut_intervals, new.cut_intervals) +# # print("MATRIX NAME:", outfile.name) +# print(set(os.listdir(ROOT + "QC/"))) +# assert are_files_equal(ROOT + "QC/QC.log", qc_folder + "/QC.log") +# assert set(os.listdir(ROOT + "QC/")) == set(os.listdir(qc_folder)) + +# # accept delta of 80 kb, file size is around 4.5 MB +# assert abs(os.path.getsize(ROOT + "hicBuildMatrixMicroC/chromosome_sizes/test.bam") - os.path.getsize(outfile_bam.name)) < delta + +# os.unlink(outfile.name) +# shutil.rmtree(qc_folder) +# # os.unlink("/tmp/test.bam") + + +# def test_build_matrix_cooler(): +# outfile = NamedTemporaryFile(suffix='.cool', delete=False) +# outfile.close() +# outfile_bam = NamedTemporaryFile(suffix='.bam', delete=False) +# outfile.close() +# qc_folder = mkdtemp(prefix="testQC_") +# args = "-s {} {} --outFileName {} -bs 5000 -b {} --QCfolder {} --threads 4 \ +# --restrictionSequence GATC --danglingSequence GATC -rs {}".format(sam_R1, sam_R2, +# outfile.name, outfile_bam.name, +# qc_folder, dpnii_file).split() +# # hicBuildMatrixMicroC.main(args) +# compute(hicBuildMatrixMicroC.main, args, 5) + +# test = hm.hiCMatrix(ROOT + "small_test_matrix_parallel.h5") +# new = hm.hiCMatrix(outfile.name) + +# nt.assert_equal(test.matrix.data, new.matrix.data) +# # nt.assert_equal(test.cut_intervals, new.cut_intervals) +# nt.assert_equal(len(new.cut_intervals), len(test.cut_intervals)) +# cut_interval_new_ = [] +# cut_interval_test_ = [] +# for x in new.cut_intervals: +# cut_interval_new_.append(x[:3]) +# for x in test.cut_intervals: +# cut_interval_test_.append(x[:3]) + +# nt.assert_equal(cut_interval_new_, cut_interval_test_) +# # print(set(os.listdir(ROOT + "QC/"))) +# assert are_files_equal(ROOT + "QC/QC.log", qc_folder + "/QC.log") +# assert set(os.listdir(ROOT + "QC/")) == set(os.listdir(qc_folder)) + +# os.unlink(outfile.name) +# shutil.rmtree(qc_folder) + + +# def test_build_matrix_cooler_metadata(): +# outfile = NamedTemporaryFile(suffix='.cool', delete=False) +# outfile.close() +# outfile_bam = NamedTemporaryFile(suffix='.bam', delete=False) +# outfile.close() +# qc_folder = mkdtemp(prefix="testQC_") +# args = "-s {} {} --outFileName {} -bs 5000 -b {} --QCfolder {} --threads 4 --genomeAssembly dm3 \ +# --restrictionSequence GATC --danglingSequence GATC -rs {}".format(sam_R1, sam_R2, +# outfile.name, outfile_bam.name, +# qc_folder, dpnii_file).split() +# # hicBuildMatrixMicroC.main(args) +# compute(hicBuildMatrixMicroC.main, args, 5) + +# test = hm.hiCMatrix(ROOT + "small_test_matrix_parallel.h5") +# new = hm.hiCMatrix(outfile.name) + +# nt.assert_equal(test.matrix.data, new.matrix.data) +# # nt.assert_equal(test.cut_intervals, new.cut_intervals) +# nt.assert_equal(len(new.cut_intervals), len(test.cut_intervals)) +# cut_interval_new_ = [] +# cut_interval_test_ = [] +# for x in new.cut_intervals: +# cut_interval_new_.append(x[:3]) +# for x in test.cut_intervals: +# cut_interval_test_.append(x[:3]) + +# nt.assert_equal(cut_interval_new_, cut_interval_test_) +# # print(set(os.listdir(ROOT + "QC/"))) +# assert are_files_equal(ROOT + "QC/QC.log", qc_folder + "/QC.log") +# assert set(os.listdir(ROOT + "QC/")) == set(os.listdir(qc_folder)) + +# outfile_metadata = NamedTemporaryFile(suffix='.txt', delete=False) +# outfile_metadata.close() +# args = "-m {} -o {}".format(outfile.name, outfile_metadata.name).split() +# hicInfo.main(args) +# assert are_files_equal(ROOT + "hicBuildMatrixMicroC/metadata.txt", outfile_metadata.name, delta=7) + +# os.unlink(outfile.name) +# shutil.rmtree(qc_folder) + + +# def test_build_matrix_cooler_multiple(): +# outfile = NamedTemporaryFile(suffix='.mcool', delete=False) +# outfile.close() +# outfile_bam = NamedTemporaryFile(suffix='.bam', delete=False) +# outfile.close() +# qc_folder = mkdtemp(prefix="testQC_") +# args = "-s {} {} --outFileName {} -bs 5000 10000 20000 -b {} --QCfolder {} --threads 4 \ +# --restrictionSequence GATC --danglingSequence GATC -rs {}".format(sam_R1, sam_R2, +# outfile.name, outfile_bam.name, +# qc_folder, dpnii_file).split() +# # hicBuildMatrixMicroC.main(args) +# compute(hicBuildMatrixMicroC.main, args, 5) + +# test_5000 = hm.hiCMatrix(ROOT + "hicBuildMatrixMicroC/multi_small_test_matrix.mcool::/resolutions/5000") +# test_10000 = hm.hiCMatrix(ROOT + "hicBuildMatrixMicroC/multi_small_test_matrix.mcool::/resolutions/10000") +# test_20000 = hm.hiCMatrix(ROOT + "hicBuildMatrixMicroC/multi_small_test_matrix.mcool::/resolutions/20000") + +# new_5000 = hm.hiCMatrix(outfile.name + '::/resolutions/5000') +# new_10000 = hm.hiCMatrix(outfile.name + '::/resolutions/10000') +# new_20000 = hm.hiCMatrix(outfile.name + '::/resolutions/20000') + +# nt.assert_equal(test_5000.matrix.data, new_5000.matrix.data) +# nt.assert_equal(test_10000.matrix.data, new_10000.matrix.data) +# nt.assert_equal(test_20000.matrix.data, new_20000.matrix.data) + +# # nt.assert_equal(test.cut_intervals, new.cut_intervals) +# nt.assert_equal(len(new_5000.cut_intervals), len(test_5000.cut_intervals)) +# nt.assert_equal(len(new_10000.cut_intervals), len(test_10000.cut_intervals)) +# nt.assert_equal(len(new_20000.cut_intervals), len(test_20000.cut_intervals)) + +# cut_interval_new_ = [] +# cut_interval_test_ = [] +# for x in new_5000.cut_intervals: +# cut_interval_new_.append(x[:3]) +# for x in test_5000.cut_intervals: +# cut_interval_test_.append(x[:3]) + +# nt.assert_equal(cut_interval_new_, cut_interval_test_) + +# cut_interval_new_ = [] +# cut_interval_test_ = [] +# for x in new_10000.cut_intervals: +# cut_interval_new_.append(x[:3]) +# for x in test_10000.cut_intervals: +# cut_interval_test_.append(x[:3]) + +# nt.assert_equal(cut_interval_new_, cut_interval_test_) + +# cut_interval_new_ = [] +# cut_interval_test_ = [] +# for x in new_20000.cut_intervals: +# cut_interval_new_.append(x[:3]) +# for x in test_20000.cut_intervals: +# cut_interval_test_.append(x[:3]) + +# nt.assert_equal(cut_interval_new_, cut_interval_test_) +# # print(set(os.listdir(ROOT + "QC/"))) +# assert are_files_equal(ROOT + "QC/QC.log", qc_folder + "/QC.log") +# assert set(os.listdir(ROOT + "QC/")) == set(os.listdir(qc_folder)) + +# os.unlink(outfile.name) +# shutil.rmtree(qc_folder) + + +# def test_build_matrix_rf(): +# outfile = NamedTemporaryFile(suffix='.h5', delete=False) +# outfile.close() +# qc_folder = mkdtemp(prefix="testQC_") +# args = "-s {} {} -rs {} --outFileName {} --QCfolder {} " \ +# "--restrictionSequence GATC " \ +# "--danglingSequence GATC " \ +# "--minDistance 150 " \ +# "--maxLibraryInsertSize 1500 --threads 4".format(sam_R1, sam_R2, dpnii_file, +# outfile.name, +# qc_folder).split() +# # hicBuildMatrixMicroC.main(args) +# compute(hicBuildMatrixMicroC.main, args, 5) + +# test = hm.hiCMatrix(ROOT + "small_test_rf_matrix.h5") +# new = hm.hiCMatrix(outfile.name) + +# nt.assert_equal(test.matrix.data, new.matrix.data) +# nt.assert_equal(test.cut_intervals, new.cut_intervals) + +# print(set(os.listdir(ROOT + "QC_rc/"))) +# assert are_files_equal(ROOT + "QC_rc/QC.log", qc_folder + "/QC.log") +# assert set(os.listdir(ROOT + "QC_rc/")) == set(os.listdir(qc_folder)) + +# os.unlink(outfile.name) +# shutil.rmtree(qc_folder) + + +# def test_build_matrix_rf_multi(): +# outfile = NamedTemporaryFile(suffix='.h5', delete=False) +# outfile.close() +# qc_folder = mkdtemp(prefix="testQC_") +# args = "-s {} {} -rs {} {} --outFileName {} --QCfolder {} " \ +# "--restrictionSequence GATC AAGCTT " \ +# "--danglingSequence GATC AGCT " \ +# "--minDistance 150 " \ +# "--maxLibraryInsertSize 1500 --threads 4".format(sam_R1, sam_R2, dpnii_file, ROOT + 'hicFindRestSite/hindIII.bed', +# outfile.name, +# qc_folder).split() +# # --danglingSequence GATC AGCT --restrictionSequence GATC AAGCTT +# # hicBuildMatrixMicroC.main(args) +# compute(hicBuildMatrixMicroC.main, args, 5) + +# test = hm.hiCMatrix(ROOT + "small_test_rf_matrix_multiple.h5") +# new = hm.hiCMatrix(outfile.name) + +# nt.assert_equal(test.matrix.data, new.matrix.data) +# nt.assert_equal(test.cut_intervals, new.cut_intervals) + +# print(set(os.listdir(ROOT + "QC_rc_multiple/"))) +# assert are_files_equal(ROOT + "QC_rc_multiple/QC.log", qc_folder + "/QC.log") +# assert set(os.listdir(ROOT + "QC_rc_multiple/")) == set(os.listdir(qc_folder)) + +# os.unlink(outfile.name) +# shutil.rmtree(qc_folder) diff --git a/setup.py b/setup.py index 841292a0..bf591a95 100755 --- a/setup.py +++ b/setup.py @@ -128,7 +128,7 @@ def checkProgramIsInstalled(self, program, args, where_to_download, author='Joachim Wolff, Leily Rabbani, Bjoern Gruening, Vivek Bhardwaj, Fidel Ramírez', author_email='deeptools@googlegroups.com', packages=find_packages(), - scripts=['bin/hicFindRestSite', 'bin/hicAggregateContacts', 'bin/hicBuildMatrix', 'bin/hicCorrectMatrix', + scripts=['bin/hicFindRestSite', 'bin/hicAggregateContacts', 'bin/hicBuildMatrix', 'bin/hicBuildMatrixMicroC', 'bin/hicCorrectMatrix', 'bin/hicCorrelate', 'bin/hicFindTADs', 'bin/hicMergeMatrixBins', 'bin/hicPlotMatrix', 'bin/hicPlotDistVsCounts', 'bin/hicPlotTADs', 'bin/hicSumMatrices', 'bin/hicInfo', 'bin/hicexplorer', 'bin/hicQC', 'bin/hicCompareMatrices', 'bin/hicPCA', 'bin/hicTransform', 'bin/hicPlotViewpoint', From 37e776b862973daddb1d3b057c7a6910fe500d00 Mon Sep 17 00:00:00 2001 From: Joachim Wolff Date: Tue, 26 Nov 2024 19:33:16 +0100 Subject: [PATCH 2/6] Making test cases for hicBuildMatrix and hicBuildMatrixMicroC run. Liniting, and updating to Python 3.11 and 3.12. Increasing the version number to 3.7.6 --- azure-pipelines.yml | 10 +- docs/content/News.rst | 8 + hicexplorer/_version.py | 2 +- hicexplorer/hicBuildMatrix.py | 23 +- hicexplorer/hicBuildMatrixMicroC.py | 16 +- hicexplorer/hicPlotDistVsCounts.py | 10 +- hicexplorer/hicPrepareQCreport.py | 9 +- hicexplorer/lib/buildMatrixMethods.py | 54 ++-- .../test/general/test_hicBuildMatrix.py | 2 +- .../test/general/test_hicBuildMatrixMicroC.py | 269 +----------------- .../test_data/QC_no_restriction_site/QC.log | 24 ++ .../QC_no_restriction_site/QC_table.txt | 2 + .../discarded_table.txt | 2 + .../QC_no_restriction_site/distance.png | Bin 0 -> 76693 bytes .../QC_no_restriction_site/distance_table.txt | 2 + .../QC_no_restriction_site/hicQC.html | 234 +++++++++++++++ .../pairs_discarded.png | Bin 0 -> 122153 bytes .../pairs_sequenced.png | Bin 0 -> 45459 bytes .../read_orientation.png | Bin 0 -> 67947 bytes .../read_orientation_table.txt | 2 + .../unmapable_table.txt | 2 + .../unmappable_and_non_unique.png | Bin 0 -> 84352 bytes .../test/test_data/number_of_tests.txt | 2 +- 23 files changed, 352 insertions(+), 321 deletions(-) create mode 100644 hicexplorer/test/test_data/QC_no_restriction_site/QC.log create mode 100644 hicexplorer/test/test_data/QC_no_restriction_site/QC_table.txt create mode 100644 hicexplorer/test/test_data/QC_no_restriction_site/discarded_table.txt create mode 100644 hicexplorer/test/test_data/QC_no_restriction_site/distance.png create mode 100644 hicexplorer/test/test_data/QC_no_restriction_site/distance_table.txt create mode 100644 hicexplorer/test/test_data/QC_no_restriction_site/hicQC.html create mode 100644 hicexplorer/test/test_data/QC_no_restriction_site/pairs_discarded.png create mode 100644 hicexplorer/test/test_data/QC_no_restriction_site/pairs_sequenced.png create mode 100644 hicexplorer/test/test_data/QC_no_restriction_site/read_orientation.png create mode 100644 hicexplorer/test/test_data/QC_no_restriction_site/read_orientation_table.txt create mode 100644 hicexplorer/test/test_data/QC_no_restriction_site/unmapable_table.txt create mode 100644 hicexplorer/test/test_data/QC_no_restriction_site/unmappable_and_non_unique.png diff --git a/azure-pipelines.yml b/azure-pipelines.yml index fc09c773..e8560053 100644 --- a/azure-pipelines.yml +++ b/azure-pipelines.yml @@ -15,7 +15,10 @@ jobs: python.version: '3.9' Python310: python.version: '3.10' - + Python311: + python.version: '3.11' + Python312: + python.version: '3.12' steps: - bash: | @@ -66,6 +69,11 @@ jobs: python.version: '3.9' Python310: python.version: '3.10' + Python311: + python.version: '3.11' + Python312: + python.version: '3.12' + steps: - bash: | diff --git a/docs/content/News.rst b/docs/content/News.rst index 03b4c1e6..861222b3 100644 --- a/docs/content/News.rst +++ b/docs/content/News.rst @@ -1,6 +1,14 @@ News and Developments ===================== +Release 3.7.6 +------------- +**27 November 2024** + +- Add a new hicBuildMatrixMicroC script to build matrices from Micro-C data. It is the same as hicBuildMatrix but without the enforcement of the restriction cut site file, restriction sequence and dangling end since this is not necessary for Micro-C data. +- Update to include support for Python 3.11 and 3.12 + + Release 3.7.5 ------------- **June 2024** diff --git a/hicexplorer/_version.py b/hicexplorer/_version.py index bbd9c98c..043cd286 100644 --- a/hicexplorer/_version.py +++ b/hicexplorer/_version.py @@ -2,4 +2,4 @@ # This file is originally generated from Git information by running 'setup.py # version'. Distribution tarballs contain a pre-generated copy of this file. -__version__ = '3.7.5' +__version__ = '3.7.6' diff --git a/hicexplorer/hicBuildMatrix.py b/hicexplorer/hicBuildMatrix.py index 2188fcfb..7ded21d2 100644 --- a/hicexplorer/hicBuildMatrix.py +++ b/hicexplorer/hicBuildMatrix.py @@ -261,21 +261,10 @@ def main(args=None): """ args = parse_arguments().parse_args(args) - + createMatrix(pOutFileName=args.outFileName, pMaxDistance=args.maxDistance, pMaxLibraryInsertSize=args.maxLibraryInsertSize, pQCfolder=args.QCfolder, - pThreads=args.threads, pDanglingSequence=args.danglingSequence, pRestrictionSequence=args.restrictionSequence, pSamFiles=args.samFiles, - pDoTestRun=args.doTestRun, pOutBam=args.outBam, pChromosomeSizes=args.chromosomeSizes, pRestrictionCutFile=args.restrictionCutFile, - pRegion=args.region, pBinSize=args.binSize, pInputBufferSize=args.inputBufferSize, pMinDistance=args.minDistance, - pDoTestRunLines=args.doTestRunLines, pSkipDuplicationCheck=args.skipDuplicationCheck, pMinMappingQuality=args.minMappingQuality, - pKeepSelfCircles=args.keepSelfCircles, pKeepSelfLigation=args.keepSelfLigation, pGenomeAssembly=args.genomeAssembly) - - -class Tester(object): - def __init__(self): - hic_test_data_dir = os.environ.get('HIC_TEST_DATA_DIR', False) - if hic_test_data_dir: - self.root = hic_test_data_dir - else: - self.root = os.path.dirname( - os.path.abspath(__file__)) + "/test/test_data/" - self.bam_file_1 = os.path.join(self.root, "hic.bam") + pThreads=args.threads, pDanglingSequence=args.danglingSequence, pRestrictionSequence=args.restrictionSequence, pSamFiles=args.samFiles, + pDoTestRun=args.doTestRun, pOutBam=args.outBam, pChromosomeSizes=args.chromosomeSizes, pRestrictionCutFile=args.restrictionCutFile, + pRegion=args.region, pBinSize=args.binSize, pInputBufferSize=args.inputBufferSize, pMinDistance=args.minDistance, + pDoTestRunLines=args.doTestRunLines, pSkipDuplicationCheck=args.skipDuplicationCheck, pMinMappingQuality=args.minMappingQuality, + pKeepSelfCircles=args.keepSelfCircles, pKeepSelfLigation=args.keepSelfLigation, pGenomeAssembly=args.genomeAssembly) diff --git a/hicexplorer/hicBuildMatrixMicroC.py b/hicexplorer/hicBuildMatrixMicroC.py index 1e8038aa..930b339e 100644 --- a/hicexplorer/hicBuildMatrixMicroC.py +++ b/hicexplorer/hicBuildMatrixMicroC.py @@ -51,7 +51,6 @@ def parse_arguments(args=None): metavar='FOLDER', required=True) - parserOpt = parser.add_argument_group('Optional arguments') parserOpt.add_argument('--outBam', '-b', @@ -77,8 +76,6 @@ def parse_arguments(args=None): nargs='+', required=True) - - parserOpt.add_argument('--maxLibraryInsertSize', help='The maximum library insert size defines different cut offs based on the maximum expected ' 'library size. *This is not the average fragment size* but the higher end of the ' @@ -104,7 +101,6 @@ def parse_arguments(args=None): type=genomicRegion ) - parserOpt.add_argument('--keepSelfCircles', help='If set, outward facing reads without any restriction fragment (self circles) are kept. ' 'They will be counted and shown in the QC plots.', @@ -210,13 +206,13 @@ def main(args=None): """ args = parse_arguments().parse_args(args) - + createMatrix(pOutFileName=args.outFileName, pMaxDistance=None, pMaxLibraryInsertSize=args.maxLibraryInsertSize, pQCfolder=args.QCfolder, - pThreads=args.threads, pDanglingSequence=None, pRestrictionSequence=None, pSamFiles=args.samFiles, - pDoTestRun=args.doTestRun, pOutBam=args.outBam, pChromosomeSizes=args.chromosomeSizes, pRestrictionCutFile=None, - pRegion=args.region, pBinSize=args.binSize, pInputBufferSize=args.inputBufferSize, pMinDistance=None, - pDoTestRunLines=args.doTestRunLines, pSkipDuplicationCheck=args.skipDuplicationCheck, pMinMappingQuality=args.minMappingQuality, - pKeepSelfCircles=args.keepSelfCircles, pKeepSelfLigation=None, pGenomeAssembly=args.genomeAssembly) + pThreads=args.threads, pDanglingSequence=None, pRestrictionSequence=None, pSamFiles=args.samFiles, + pDoTestRun=args.doTestRun, pOutBam=args.outBam, pChromosomeSizes=args.chromosomeSizes, pRestrictionCutFile=None, + pRegion=args.region, pBinSize=args.binSize, pInputBufferSize=args.inputBufferSize, pMinDistance=None, + pDoTestRunLines=args.doTestRunLines, pSkipDuplicationCheck=args.skipDuplicationCheck, pMinMappingQuality=args.minMappingQuality, + pKeepSelfCircles=args.keepSelfCircles, pKeepSelfLigation=None, pGenomeAssembly=args.genomeAssembly) class Tester(object): diff --git a/hicexplorer/hicPlotDistVsCounts.py b/hicexplorer/hicPlotDistVsCounts.py index b147bd93..4a441641 100644 --- a/hicexplorer/hicPlotDistVsCounts.py +++ b/hicexplorer/hicPlotDistVsCounts.py @@ -156,20 +156,20 @@ def compute_distance_mean(hicmat, maxdepth=None, perchr=False, custom_cut_interv >>> hic.matrix = csr_matrix(matrix) >>> hic.setMatrix(hic.matrix, cut_intervals) >>> compute_distance_mean(hic) - {'all': OrderedDict([(0, (3.0, 7)), (10, (6.0, 5)), (20, (10.0, 3)), (30, (3.0, 1))])} + {'all': OrderedDict({0: (3.0, 7), 10: (6.0, 5), 20: (10.0, 3), 30: (3.0, 1)})} >>> compute_distance_mean(hic, perchr=True) - {'a': OrderedDict([(0, (1.25, 4)), (10, (10.0, 3)), (20, (5.0, 2)), (30, (3.0, 1))]), 'b': OrderedDict([(0, (5.333333333333333, 3)), (10, (0.0, 2)), (20, (20.0, 1))])} + {'a': OrderedDict({0: (1.25, 4), 10: (10.0, 3), 20: (5.0, 2), 30: (3.0, 1)}), 'b': OrderedDict({0: (5.333333333333333, 3), 10: (0.0, 2), 20: (20.0, 1)})} >>> custom_cut = [('tad1', 0, 10, 1), ('tad1', 10, 20, 1), ('tad2', 0, 10, 1), ... ('tad2', 10, 20, 1), ('tad3', 0, 10, 1), ('tad3', 10, 20, 1), ('tad3', 20, 30, 1)] >>> compute_distance_mean(hic, custom_cut_intervals=custom_cut) - {'all': OrderedDict([(0, (3.0, 7)), (10, (3.75, 4)), (20, (20.0, 1))])} + {'all': OrderedDict({0: (3.0, 7), 10: (3.75, 4), 20: (20.0, 1)})} >>> compute_distance_mean(hic, perchr=True, custom_cut_intervals=custom_cut) - {'a': OrderedDict([(0, (1.25, 4)), (10, (7.5, 2))]), 'b': OrderedDict([(0, (5.333333333333333, 3)), (10, (0.0, 2)), (20, (20.0, 1))])} + {'a': OrderedDict({0: (1.25, 4), 10: (7.5, 2)}), 'b': OrderedDict({0: (5.333333333333333, 3), 10: (0.0, 2), 20: (20.0, 1)})} >>> custom_cut = [('_ignore_0', 0, 10, 1), ('0', 0, 10, 1), ... ('0', 10, 20, 1), ('_ignore_3', 0, 10, 1), ... ('1', 0, 10, 1), ('1', 10, 20, 1), ('1', 20, 30, 1)] >>> compute_distance_mean(hic, custom_cut_intervals=custom_cut) - {'all': OrderedDict([(0, (4.0, 5)), (10, (5.0, 3)), (20, (20.0, 1))])} + {'all': OrderedDict({0: (4.0, 5), 10: (5.0, 3), 20: (20.0, 1)})} # >>> compute_distance_mean(hic, custom_cut_intervals=custom_cut, perchr=True) # {'a': OrderedDict([(0, (2.0, 2)), (10, (15.0, 1))]), 'b': OrderedDict([(0, (5.333333333333333, 3)), (10, (0.0, 2)), (20, (20.0, 1))])} diff --git a/hicexplorer/hicPrepareQCreport.py b/hicexplorer/hicPrepareQCreport.py index 2afc267e..d59d390c 100644 --- a/hicexplorer/hicPrepareQCreport.py +++ b/hicexplorer/hicPrepareQCreport.py @@ -72,7 +72,12 @@ def save_html(filename, unmap_table, discard_table, distance_table, orientation_ html_content = html_content.replace("%%TABLE_ORIENTATION%%", orientation_table.style .format(lambda x: '{:,}'.format(x) if x > 1 else '{:.2%}'.format(x)).to_html(classes='df')) - all_table = all_table.drop(['Min rest. site distance', 'Max library insert size'], axis=1) + if 'Min rest. site distance' in all_table.columns: + all_table = all_table.drop(['Min rest. site distance'], axis=1) + + if 'Max library insert size' in all_table.columns: + all_table = all_table.drop(['Max library insert size'], axis=1) + # all_table = all_table.drop(['Min rest. site distance', 'Max library insert size'], axis=1) html_content = html_content.replace("%%TABLE%%", all_table.style.to_html(classes='df')) with open(filename, 'w') as fh: @@ -268,6 +273,8 @@ def main(args=None): in_log_part = False log.debug('Processing {}\n'.format(fh.name)) for line in fh.readlines(): + log.debug('line {}'.format(line)) + line = line.strip() if line.startswith("File"): in_log_part = True if in_log_part is True: diff --git a/hicexplorer/lib/buildMatrixMethods.py b/hicexplorer/lib/buildMatrixMethods.py index 5f7e7628..fefe2c4e 100644 --- a/hicexplorer/lib/buildMatrixMethods.py +++ b/hicexplorer/lib/buildMatrixMethods.py @@ -7,6 +7,7 @@ import time from os import unlink import os +from pathlib import Path from io import StringIO import traceback import warnings @@ -37,6 +38,7 @@ import logging log = logging.getLogger(__name__) + class C_Interval(Structure): """Struct to map a Interval form intervaltree as a multiprocessing.sharedctype""" _fields_ = [("begin", c_uint), @@ -85,6 +87,7 @@ def is_duplicated(self, chrom1, start1, chrom2, start2): self.pos_matrix.add(id_string) return False + def intervalListToIntervalTree(interval_list): r""" given a dictionary containing tuples of chrom, start, end, @@ -554,7 +557,6 @@ def readBamFiles(pFileOneIterator, pFileTwoIterator, pNumberOfItemsPerBuffer, pS return buffer_mate1, buffer_mate2, False, duplicated_pairs, one_mate_unmapped, one_mate_not_unique, one_mate_low_quality, iter_num - len(buffer_mate1) - def process_data(pMateBuffer1, pMateBuffer2, pMinMappingQuality, pKeepSelfCircles, pRestrictionSequence, pKeepSelfLigation, pMatrixSize, pRfPositions, pRefId2name, @@ -866,13 +868,14 @@ def process_data(pMateBuffer1, pMateBuffer2, pMinMappingQuality, return return + def createMatrix(pOutFileName, pMaxDistance, pMaxLibraryInsertSize, pQCfolder, pThreads, pDanglingSequence, pRestrictionSequence, pSamFiles, pDoTestRun, pOutBam, pChromosomeSizes, pRestrictionCutFile, pRegion, pBinSize, pInputBufferSize, - pDoTestRunLines, pSkipDuplicationCheck, pMinMappingQuality, + pDoTestRunLines, pSkipDuplicationCheck, pMinMappingQuality, pKeepSelfCircles, pKeepSelfLigation, pMinDistance, pGenomeAssembly): - # pOutFileName.name = pOutFileName.name.strip() + # pOutFileName.name = pOutFileName.name.strip() # log.debug('pOutFileName.name: {}'.format(pOutFileName.name.endswith('.h5'))) if not pOutFileName.name.endswith('.h5') and not pOutFileName.name.endswith('.cool'): if '.mcool' not in pOutFileName.name: @@ -1262,19 +1265,19 @@ def createMatrix(pOutFileName, pMaxDistance, pMaxLibraryInsertSize, pQCfolder, if pMinDistance: intermediate_qc_log.write(""" - File\t{}\t\t - Sequenced reads\t{}\t\t - Min rest. site distance\t{}\t\t - Max library insert size\t{}\t\t +File\t{}\t\t +Sequenced reads\t{}\t\t +Min rest. site distance\t{}\t\t +Max library insert size\t{}\t\t - """.format(pOutFileName.name, iter_num, pMinDistance, pMaxLibraryInsertSize)) +""".format(pOutFileName.name, iter_num, pMinDistance, pMaxLibraryInsertSize)) else: intermediate_qc_log.write(""" - File\t{}\t\t - Sequenced reads\t{}\t\t - Max library insert size\t{}\t\t +File\t{}\t\t +Sequenced reads\t{}\t\t +Max library insert size\t{}\t\t - """.format(pOutFileName.name, iter_num, pMaxLibraryInsertSize)) +""".format(pOutFileName.name, iter_num, pMaxLibraryInsertSize)) intermediate_qc_log.write( "#\tcount\t(percentage w.r.t. total sequenced reads)\n") @@ -1307,20 +1310,20 @@ def createMatrix(pOutFileName, pMaxDistance, pMaxLibraryInsertSize, pQCfolder, # dangling_sequences[pRestrictionSequence[i]]['pat_forw'] intermediate_qc_log.write("dangling end {} (restriction sequence {})\t{}\t({:.2f})\n". format(dangling_sequences[key]['pat_forw'], key, dangling_end[key], 100 * float(dangling_end[key]) / mappable_unique_high_quality_pairs)) - else: - intermediate_qc_log.write("dangling end\t{}\t({:.2f})\n". - format(0, 100 * float(0) / mappable_unique_high_quality_pairs)) + # else: + # intermediate_qc_log.write("dangling end\t{}\t({:.2f})\n". + # format(0, 100 * float(0) / mappable_unique_high_quality_pairs)) if pRestrictionCutFile is not None: intermediate_qc_log.write("self ligation{}\t{}\t({:.2f})\n". - format(msg, self_ligation, 100 * float(self_ligation) / mappable_unique_high_quality_pairs)) + format(msg, self_ligation, 100 * float(self_ligation) / mappable_unique_high_quality_pairs)) intermediate_qc_log.write("One mate not close to rest site\t{}\t({:.2f})\n". - format(mate_not_close_to_rf, 100 * float(mate_not_close_to_rf) / mappable_unique_high_quality_pairs)) - intermediate_qc_log.write("self circle\t{}\t({:.2f})\n". - format(self_circle, 100 * float(self_circle) / mappable_unique_high_quality_pairs)) - + format(mate_not_close_to_rf, 100 * float(mate_not_close_to_rf) / mappable_unique_high_quality_pairs)) intermediate_qc_log.write("same fragment\t{}\t({:.2f})\n". format(same_fragment, 100 * float(same_fragment) / mappable_unique_high_quality_pairs)) + if pRestrictionCutFile is not None: + intermediate_qc_log.write("self circle\t{}\t({:.2f})\n". + format(self_circle, 100 * float(self_circle) / mappable_unique_high_quality_pairs)) intermediate_qc_log.write("duplicated pairs\t{}\t({:.2f})\n". format(duplicated_pairs, 100 * float(duplicated_pairs) / mappable_unique_high_quality_pairs)) @@ -1403,3 +1406,14 @@ def createMatrix(pOutFileName, pMaxDistance, pMaxLibraryInsertSize, pQCfolder, else: if not pDoTestRun: hic_ma.save(pOutFileName.name, pHiCInfo=hic_metadata) + + +class Tester(object): + def __init__(self): + hic_test_data_dir = os.environ.get('HIC_TEST_DATA_DIR', False) + if hic_test_data_dir: + self.root = hic_test_data_dir + else: + self.root = os.path.dirname(os.path.dirname( + os.path.abspath(__file__))) + "/test/test_data/" + self.bam_file_1 = os.path.join(self.root, "hic.bam") diff --git a/hicexplorer/test/general/test_hicBuildMatrix.py b/hicexplorer/test/general/test_hicBuildMatrix.py index cd869360..e7bf666b 100644 --- a/hicexplorer/test/general/test_hicBuildMatrix.py +++ b/hicexplorer/test/general/test_hicBuildMatrix.py @@ -55,7 +55,7 @@ def test_build_matrix(capsys): nt.assert_equal(test.cut_intervals, new.cut_intervals) # print("MATRIX NAME:", outfile.name) print(set(os.listdir(ROOT + "QC/"))) - assert are_files_equal(ROOT + "QC/QC.log", qc_folder + "/QC.log") + assert are_files_equal(ROOT + "QC/QC.log", qc_folder + "/QC.log", delta=2) assert set(os.listdir(ROOT + "QC/")) == set(os.listdir(qc_folder)) # accept delta of 80 kb, file size is around 4.5 MB diff --git a/hicexplorer/test/general/test_hicBuildMatrixMicroC.py b/hicexplorer/test/general/test_hicBuildMatrixMicroC.py index c1fdbd0f..0213375e 100644 --- a/hicexplorer/test/general/test_hicBuildMatrixMicroC.py +++ b/hicexplorer/test/general/test_hicBuildMatrixMicroC.py @@ -43,279 +43,20 @@ def test_build_matrix(capsys): outfile.close() qc_folder = mkdtemp(prefix="testQC_") args = "-s {} {} --outFileName {} -bs 5000 -b {} --QCfolder {} --threads 4".format(sam_R1, sam_R2, - outfile.name, outfile_bam.name, - qc_folder).split() - # hicBuildMatrixMicroC.main(args) + outfile.name, outfile_bam.name, + qc_folder).split() compute(hicBuildMatrixMicroC.main, args, 5) test = hm.hiCMatrix(ROOT + "small_test_matrix_parallel_one_rc.h5") new = hm.hiCMatrix(outfile.name) nt.assert_equal(test.matrix.data, new.matrix.data) nt.assert_equal(test.cut_intervals, new.cut_intervals) # print("MATRIX NAME:", outfile.name) - print(set(os.listdir(ROOT + "QC/"))) - assert are_files_equal(ROOT + "QC/QC.log", qc_folder + "/QC.log") - assert set(os.listdir(ROOT + "QC/")) == set(os.listdir(qc_folder)) + print(set(os.listdir(ROOT + "QC_no_restriction_site/"))) + assert are_files_equal(ROOT + "QC_no_restriction_site/QC.log", qc_folder + "/QC.log") + assert set(os.listdir(ROOT + "QC_no_restriction_site/")) == set(os.listdir(qc_folder)) # accept delta of 80 kb, file size is around 4.5 MB assert abs(os.path.getsize(ROOT + "small_test_matrix_result.bam") - os.path.getsize(outfile_bam.name)) < delta os.unlink(outfile.name) shutil.rmtree(qc_folder) - # os.unlink("/tmp/test.bam") - - -# def test_build_matrix_restriction_enzyme_region(capsys): -# outfile = NamedTemporaryFile(suffix='.h5', delete=False) -# outfile.close() -# outfile_bam = NamedTemporaryFile(suffix='.bam', delete=False) -# outfile.close() -# qc_folder = mkdtemp(prefix="testQC_") -# args = "-s {} {} --outFileName {} -bs 5000 -b {} --QCfolder {} --threads 4 --region {}".format(sam_R1, sam_R2, outfile.name, outfile_bam.name, qc_folder, 'chr3R').split() -# # hicBuildMatrixMicroC.main(args) -# compute(hicBuildMatrixMicroC.main, args, 5) - -# test = hm.hiCMatrix(ROOT + "small_test_matrix_parallel_two_rc_chr3R.h5") -# new = hm.hiCMatrix(outfile.name) - -# # print('test.cut_intervals {}'.format(test.cut_intervals)) -# # print('new.cut_intervals {}'.format(new.cut_intervals)) -# nt.assert_equal(test.matrix.data, new.matrix.data) -# nt.assert_equal(len(test.cut_intervals), len(new.cut_intervals)) - -# # print("MATRIX NAME:", outfile.name) -# print(set(os.listdir(ROOT + "QC_region/"))) -# assert are_files_equal(ROOT + "QC_region/QC.log", qc_folder + "/QC.log") -# assert set(os.listdir(ROOT + "QC_region/")) == set(os.listdir(qc_folder)) - -# # accept delta of 80 kb, file size is around 4.5 MB -# assert abs(os.path.getsize(ROOT + "build_region.bam") - os.path.getsize(outfile_bam.name)) < delta - -# os.unlink(outfile.name) -# shutil.rmtree(qc_folder) - - -# def test_build_matrix_chromosome_sizes(capsys): -# outfile = NamedTemporaryFile(suffix='.h5', delete=False) -# outfile.close() -# outfile_bam = NamedTemporaryFile(suffix='.bam', delete=False) -# outfile.close() -# qc_folder = mkdtemp(prefix="testQC_") -# args = "-s {} {} --outFileName {} -bs 5000 -b {} --QCfolder {} --threads 4 --chromosomeSizes {} \ -# --restrictionSequence GATC --danglingSequence GATC -rs {}".format(sam_R1, sam_R2, -# outfile.name, outfile_bam.name, -# qc_folder, ROOT + 'hicBuildMatrixMicroC/dm3.chrom.sizes', dpnii_file).split() -# # hicBuildMatrixMicroC.main(args) -# compute(hicBuildMatrixMicroC.main, args, 5) - -# test = hm.hiCMatrix(ROOT + "hicBuildMatrixMicroC/chromosome_sizes/small_test_chromosome_size.h5") -# new = hm.hiCMatrix(outfile.name) -# nt.assert_equal(test.matrix.data, new.matrix.data) -# nt.assert_equal(test.cut_intervals, new.cut_intervals) -# # print("MATRIX NAME:", outfile.name) -# print(set(os.listdir(ROOT + "QC/"))) -# assert are_files_equal(ROOT + "QC/QC.log", qc_folder + "/QC.log") -# assert set(os.listdir(ROOT + "QC/")) == set(os.listdir(qc_folder)) - -# # accept delta of 80 kb, file size is around 4.5 MB -# assert abs(os.path.getsize(ROOT + "hicBuildMatrixMicroC/chromosome_sizes/test.bam") - os.path.getsize(outfile_bam.name)) < delta - -# os.unlink(outfile.name) -# shutil.rmtree(qc_folder) -# # os.unlink("/tmp/test.bam") - - -# def test_build_matrix_cooler(): -# outfile = NamedTemporaryFile(suffix='.cool', delete=False) -# outfile.close() -# outfile_bam = NamedTemporaryFile(suffix='.bam', delete=False) -# outfile.close() -# qc_folder = mkdtemp(prefix="testQC_") -# args = "-s {} {} --outFileName {} -bs 5000 -b {} --QCfolder {} --threads 4 \ -# --restrictionSequence GATC --danglingSequence GATC -rs {}".format(sam_R1, sam_R2, -# outfile.name, outfile_bam.name, -# qc_folder, dpnii_file).split() -# # hicBuildMatrixMicroC.main(args) -# compute(hicBuildMatrixMicroC.main, args, 5) - -# test = hm.hiCMatrix(ROOT + "small_test_matrix_parallel.h5") -# new = hm.hiCMatrix(outfile.name) - -# nt.assert_equal(test.matrix.data, new.matrix.data) -# # nt.assert_equal(test.cut_intervals, new.cut_intervals) -# nt.assert_equal(len(new.cut_intervals), len(test.cut_intervals)) -# cut_interval_new_ = [] -# cut_interval_test_ = [] -# for x in new.cut_intervals: -# cut_interval_new_.append(x[:3]) -# for x in test.cut_intervals: -# cut_interval_test_.append(x[:3]) - -# nt.assert_equal(cut_interval_new_, cut_interval_test_) -# # print(set(os.listdir(ROOT + "QC/"))) -# assert are_files_equal(ROOT + "QC/QC.log", qc_folder + "/QC.log") -# assert set(os.listdir(ROOT + "QC/")) == set(os.listdir(qc_folder)) - -# os.unlink(outfile.name) -# shutil.rmtree(qc_folder) - - -# def test_build_matrix_cooler_metadata(): -# outfile = NamedTemporaryFile(suffix='.cool', delete=False) -# outfile.close() -# outfile_bam = NamedTemporaryFile(suffix='.bam', delete=False) -# outfile.close() -# qc_folder = mkdtemp(prefix="testQC_") -# args = "-s {} {} --outFileName {} -bs 5000 -b {} --QCfolder {} --threads 4 --genomeAssembly dm3 \ -# --restrictionSequence GATC --danglingSequence GATC -rs {}".format(sam_R1, sam_R2, -# outfile.name, outfile_bam.name, -# qc_folder, dpnii_file).split() -# # hicBuildMatrixMicroC.main(args) -# compute(hicBuildMatrixMicroC.main, args, 5) - -# test = hm.hiCMatrix(ROOT + "small_test_matrix_parallel.h5") -# new = hm.hiCMatrix(outfile.name) - -# nt.assert_equal(test.matrix.data, new.matrix.data) -# # nt.assert_equal(test.cut_intervals, new.cut_intervals) -# nt.assert_equal(len(new.cut_intervals), len(test.cut_intervals)) -# cut_interval_new_ = [] -# cut_interval_test_ = [] -# for x in new.cut_intervals: -# cut_interval_new_.append(x[:3]) -# for x in test.cut_intervals: -# cut_interval_test_.append(x[:3]) - -# nt.assert_equal(cut_interval_new_, cut_interval_test_) -# # print(set(os.listdir(ROOT + "QC/"))) -# assert are_files_equal(ROOT + "QC/QC.log", qc_folder + "/QC.log") -# assert set(os.listdir(ROOT + "QC/")) == set(os.listdir(qc_folder)) - -# outfile_metadata = NamedTemporaryFile(suffix='.txt', delete=False) -# outfile_metadata.close() -# args = "-m {} -o {}".format(outfile.name, outfile_metadata.name).split() -# hicInfo.main(args) -# assert are_files_equal(ROOT + "hicBuildMatrixMicroC/metadata.txt", outfile_metadata.name, delta=7) - -# os.unlink(outfile.name) -# shutil.rmtree(qc_folder) - - -# def test_build_matrix_cooler_multiple(): -# outfile = NamedTemporaryFile(suffix='.mcool', delete=False) -# outfile.close() -# outfile_bam = NamedTemporaryFile(suffix='.bam', delete=False) -# outfile.close() -# qc_folder = mkdtemp(prefix="testQC_") -# args = "-s {} {} --outFileName {} -bs 5000 10000 20000 -b {} --QCfolder {} --threads 4 \ -# --restrictionSequence GATC --danglingSequence GATC -rs {}".format(sam_R1, sam_R2, -# outfile.name, outfile_bam.name, -# qc_folder, dpnii_file).split() -# # hicBuildMatrixMicroC.main(args) -# compute(hicBuildMatrixMicroC.main, args, 5) - -# test_5000 = hm.hiCMatrix(ROOT + "hicBuildMatrixMicroC/multi_small_test_matrix.mcool::/resolutions/5000") -# test_10000 = hm.hiCMatrix(ROOT + "hicBuildMatrixMicroC/multi_small_test_matrix.mcool::/resolutions/10000") -# test_20000 = hm.hiCMatrix(ROOT + "hicBuildMatrixMicroC/multi_small_test_matrix.mcool::/resolutions/20000") - -# new_5000 = hm.hiCMatrix(outfile.name + '::/resolutions/5000') -# new_10000 = hm.hiCMatrix(outfile.name + '::/resolutions/10000') -# new_20000 = hm.hiCMatrix(outfile.name + '::/resolutions/20000') - -# nt.assert_equal(test_5000.matrix.data, new_5000.matrix.data) -# nt.assert_equal(test_10000.matrix.data, new_10000.matrix.data) -# nt.assert_equal(test_20000.matrix.data, new_20000.matrix.data) - -# # nt.assert_equal(test.cut_intervals, new.cut_intervals) -# nt.assert_equal(len(new_5000.cut_intervals), len(test_5000.cut_intervals)) -# nt.assert_equal(len(new_10000.cut_intervals), len(test_10000.cut_intervals)) -# nt.assert_equal(len(new_20000.cut_intervals), len(test_20000.cut_intervals)) - -# cut_interval_new_ = [] -# cut_interval_test_ = [] -# for x in new_5000.cut_intervals: -# cut_interval_new_.append(x[:3]) -# for x in test_5000.cut_intervals: -# cut_interval_test_.append(x[:3]) - -# nt.assert_equal(cut_interval_new_, cut_interval_test_) - -# cut_interval_new_ = [] -# cut_interval_test_ = [] -# for x in new_10000.cut_intervals: -# cut_interval_new_.append(x[:3]) -# for x in test_10000.cut_intervals: -# cut_interval_test_.append(x[:3]) - -# nt.assert_equal(cut_interval_new_, cut_interval_test_) - -# cut_interval_new_ = [] -# cut_interval_test_ = [] -# for x in new_20000.cut_intervals: -# cut_interval_new_.append(x[:3]) -# for x in test_20000.cut_intervals: -# cut_interval_test_.append(x[:3]) - -# nt.assert_equal(cut_interval_new_, cut_interval_test_) -# # print(set(os.listdir(ROOT + "QC/"))) -# assert are_files_equal(ROOT + "QC/QC.log", qc_folder + "/QC.log") -# assert set(os.listdir(ROOT + "QC/")) == set(os.listdir(qc_folder)) - -# os.unlink(outfile.name) -# shutil.rmtree(qc_folder) - - -# def test_build_matrix_rf(): -# outfile = NamedTemporaryFile(suffix='.h5', delete=False) -# outfile.close() -# qc_folder = mkdtemp(prefix="testQC_") -# args = "-s {} {} -rs {} --outFileName {} --QCfolder {} " \ -# "--restrictionSequence GATC " \ -# "--danglingSequence GATC " \ -# "--minDistance 150 " \ -# "--maxLibraryInsertSize 1500 --threads 4".format(sam_R1, sam_R2, dpnii_file, -# outfile.name, -# qc_folder).split() -# # hicBuildMatrixMicroC.main(args) -# compute(hicBuildMatrixMicroC.main, args, 5) - -# test = hm.hiCMatrix(ROOT + "small_test_rf_matrix.h5") -# new = hm.hiCMatrix(outfile.name) - -# nt.assert_equal(test.matrix.data, new.matrix.data) -# nt.assert_equal(test.cut_intervals, new.cut_intervals) - -# print(set(os.listdir(ROOT + "QC_rc/"))) -# assert are_files_equal(ROOT + "QC_rc/QC.log", qc_folder + "/QC.log") -# assert set(os.listdir(ROOT + "QC_rc/")) == set(os.listdir(qc_folder)) - -# os.unlink(outfile.name) -# shutil.rmtree(qc_folder) - - -# def test_build_matrix_rf_multi(): -# outfile = NamedTemporaryFile(suffix='.h5', delete=False) -# outfile.close() -# qc_folder = mkdtemp(prefix="testQC_") -# args = "-s {} {} -rs {} {} --outFileName {} --QCfolder {} " \ -# "--restrictionSequence GATC AAGCTT " \ -# "--danglingSequence GATC AGCT " \ -# "--minDistance 150 " \ -# "--maxLibraryInsertSize 1500 --threads 4".format(sam_R1, sam_R2, dpnii_file, ROOT + 'hicFindRestSite/hindIII.bed', -# outfile.name, -# qc_folder).split() -# # --danglingSequence GATC AGCT --restrictionSequence GATC AAGCTT -# # hicBuildMatrixMicroC.main(args) -# compute(hicBuildMatrixMicroC.main, args, 5) - -# test = hm.hiCMatrix(ROOT + "small_test_rf_matrix_multiple.h5") -# new = hm.hiCMatrix(outfile.name) - -# nt.assert_equal(test.matrix.data, new.matrix.data) -# nt.assert_equal(test.cut_intervals, new.cut_intervals) - -# print(set(os.listdir(ROOT + "QC_rc_multiple/"))) -# assert are_files_equal(ROOT + "QC_rc_multiple/QC.log", qc_folder + "/QC.log") -# assert set(os.listdir(ROOT + "QC_rc_multiple/")) == set(os.listdir(qc_folder)) - -# os.unlink(outfile.name) -# shutil.rmtree(qc_folder) diff --git a/hicexplorer/test/test_data/QC_no_restriction_site/QC.log b/hicexplorer/test/test_data/QC_no_restriction_site/QC.log new file mode 100644 index 00000000..3d5310de --- /dev/null +++ b/hicexplorer/test/test_data/QC_no_restriction_site/QC.log @@ -0,0 +1,24 @@ + +File /tmp/tmpjoshwi_n.h5 +Sequenced reads 99983 +Max library insert size 1000 + +# count (percentage w.r.t. total sequenced reads) +Pairs mappable, unique and high quality 52726 (52.73) +Hi-C contacts 37321 (37.33) +One mate unmapped 8777 (8.78) +One mate not unique 3603 (3.60) +Low mapping quality 34877 (34.88) + +# count (percentage w.r.t. mappable, unique and high quality pairs) +same fragment 15393 (29.19) +duplicated pairs 12 (0.02) + +# count (percentage w.r.t. total valid pairs used) +inter chromosomal 5955 (15.96) +Intra short range (< 20kb) 8853 (23.72) +Intra long range (>= 20kb) 22513 (60.32) +Read pair type: inward pairs 7145 (19.14) +Read pair type: outward pairs 9731 (26.07) +Read pair type: left pairs 7156 (19.17) +Read pair type: right pairs 7334 (19.65) diff --git a/hicexplorer/test/test_data/QC_no_restriction_site/QC_table.txt b/hicexplorer/test/test_data/QC_no_restriction_site/QC_table.txt new file mode 100644 index 00000000..dbaf4fdb --- /dev/null +++ b/hicexplorer/test/test_data/QC_no_restriction_site/QC_table.txt @@ -0,0 +1,2 @@ +File Sequenced reads Min rest. site distance Max library insert size Pairs mappable, unique and high quality Hi-C contacts One mate unmapped One mate not unique Low mapping quality dangling end GATC self ligation (removed) One mate not close to rest site same fragment self circle duplicated pairs inter chromosomal Intra short range (< 20kb) Intra long range (>= 20kb) Read pair type: inward pairs Read pair type: outward pairs Read pair type: left pairs Read pair type: right pairs +/tmp/tmphfn27ves.h5 99983 300 1000 52726 37321 8777 3603 34877 54 5056 0 10283 651 12 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zAi8;|4_a~9(0Sqh%hEfdTVV)Xl$11C;Yw@OSu=2sAUT^oOzd` z9pXiXn=9YFn?o&X@!?PLg(G;;X^2_?0z9+)-wXy?C?!>e9a`epz^|&$AAe9DSN21% z^o4JJ^K#>_GuaI-M= 20kb) Intra long range (>= 20kb) % +/tmp/tmphfn27ves.h5 5955 0.15956164089922564 8853 0.23721229334690924 22513 0.6032260657538652 diff --git a/hicexplorer/test/test_data/QC_no_restriction_site/hicQC.html b/hicexplorer/test/test_data/QC_no_restriction_site/hicQC.html new file mode 100644 index 00000000..0f3393aa --- /dev/null +++ b/hicexplorer/test/test_data/QC_no_restriction_site/hicQC.html @@ -0,0 +1,234 @@ + + + + + HiCExplorer QC report + + + + + + + +
+
+
+

HiCExplorer QC report

+
+ +

Pairs sequenced

+ Pairs sequenced + +

Unmappable and non-unique pairs

+ Unmappable and non-unique + +

This figure contains the fraction of reads, with respect to the total number of reads, + that did not map, that have a low quality score or that didn't map uniquely to the genome. + +

Unmappable and non-unique table

+ + + + + + + + + + + + +
Hi-C contacts Hi-C contacts_% Low mapping quality Low mapping quality_% One mate not unique One mate not unique_% One mate unmapped One mate unmapped_%
File
/tmp/tmphfn27ves.h537,32137.33%34,87734.88%3,6033.60%8,7778.78%
+ Download table (tab separated) + +

Pairs discarded

+ Pairs discarded + +

This figure contains the fraction read pairs (with respect to mappable reads) that were discarded + when building the Hi-C matrix.

+
+
Dangling ends
+
These are reads that start with the restriction site and constitute reads that were digested but no ligated.
+
Same fragment
+
These are read mates, facing inward, separated by up to 800 bp that do not have a restriction enzyme in + between. These read pairs are not valid Hi-C pairs
+
Self circle
+
self circles are defined as pairs within 25kb with 'outward' read orientation
+
Self ligation
+
These are read pairs with a restriction site in between that are within 800 bp.
+
+ +

Discarded pairs table

+ + + + + + + + + + + + + + + +
One mate not close to rest site One mate not close to rest site % dangling end GATC % duplicated pairs duplicated pairs % same fragment same fragment % self circle self circle % self ligation (removed) self ligation (removed) %
File
/tmp/tmphfn27ves.h50.00%0.00%0.10%120.02%10,28319.50%6511.23%5,0569.59%
+ Download table (tab separated) + +

Contact distance

+ Contact distance + +

Contact distance table

+ + + + + + + + + + +
inter chromosomal inter chromosomal % Intra short range (< 20kb) Intra short range (< 20kb) % Intra long range (>= 20kb) Intra long range (>= 20kb) %
File
/tmp/tmphfn27ves.h55,95515.96%8,85323.72%22,51360.32%
+ Download table (tab separated) + +

Read orientation

+ Contact distance + +

Read orientation table

+ + + + + + + + + + + + +
Read pair type: inward pairs Read pair type: inward pairs % Read pair type: outward pairs Read pair type: outward pairs % Read pair type: left pairs Read pair type: left pairs % Read pair type: right pairs Read pair type: right pairs %
File
/tmp/tmphfn27ves.h57,14522.78%9,73131.02%7,15622.81%7,33423.38%
+ Download table (tab separated) + +

Number of reads table

+ + + + + + + + + + + + + + + + + + + + + + + +
Sequenced reads Pairs mappable, unique and high quality Hi-C contacts One mate unmapped One mate not unique Low mapping quality dangling end GATC self ligation (removed) One mate not close to rest site same fragment self circle duplicated pairs inter chromosomal Intra short range (< 20kb) Intra long range (>= 20kb) Read pair type: inward pairs Read pair type: outward pairs Read pair type: left pairs Read pair type: right pairs
File
/tmp/tmphfn27ves.h599983527263732187773603348775450560102836511259558853225137145973171567334
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+ + diff --git a/hicexplorer/test/test_data/QC_no_restriction_site/pairs_discarded.png b/hicexplorer/test/test_data/QC_no_restriction_site/pairs_discarded.png new file mode 100644 index 0000000000000000000000000000000000000000..164d803fc2b1a8817750cb88b6c379ecd707f0bb GIT binary patch literal 122153 zcmeFZXH->Lmo`e3rB)f=G8QUgI3z()5Rhyr#e*ct8I>eiat>9Np;SN*NREmkSwM0! zlR?QyQc1#5keuN@i-XqNJ-*SSyT|wA-Z3uY?P`I;-fOS5=A2KMYyYhvC$)CfwpC0_ zOlzs9e^+E;S~1AP^wS?dufR_Z+bw&6|M!#aajNpq_;UN%;5xql<$;!yi@w|;8lf`+vOXgN~<|Y?*IT+g5npj!#A2@V?pJ&%aJG)CF zyuAPX{(zN@F|QZXq2HO9b}>W2$UyA-~-@VLN|1u2z z^A}A`-M32YRQ9W(HZ8kHY;Fu6@1a(yo)rx)-o>>!rRDVRuk2#~OkQ{K#`TMRaz8BR zEjgK!?&Rq?sn2q!Th@ed_}bjJi^m2 zemyPrcVzraVbA}Y{Z#hU#G09+TgebpY#&pH>rsk{q**Jp+3`7qjU>)#-sZG zY}I~@x!Jil+{}kVHpIWpv7=yTE%}`}bGmVLa&i+vimbC{2J&Lv zXU4~Z$bLCjKGW49$#hQ3phbppLC#;xME~di>TkQn%y0Ad0-hOqB1i{m%vScQF}-m#lguk*Ecd9ZOr@bcAL5=W*zX1)Yo+v&pTifLz0nfv-; zeQ%wTd}W$px!@7YMjgk>aL4vw3HNF9ORZg{ezs*?u?FLxo9X$5#XDWQ9=E 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a2mcv=AH0{$@&q^$4+NgBelF{r5}E*Lg*k2j literal 0 HcmV?d00001 diff --git a/hicexplorer/test/test_data/number_of_tests.txt b/hicexplorer/test/test_data/number_of_tests.txt index e88ff725..a7ff3ab0 100644 --- a/hicexplorer/test/test_data/number_of_tests.txt +++ b/hicexplorer/test/test_data/number_of_tests.txt @@ -1 +1 @@ -602 \ No newline at end of file +605 \ No newline at end of file From 5648c5f5ef4ed2cab69fc64057a59e6232603a04 Mon Sep 17 00:00:00 2001 From: Joachim Wolff Date: Tue, 26 Nov 2024 19:40:58 +0100 Subject: [PATCH 3/6] Add bin for hicBuildMatrixMicroC --- bin/hicBuildMatrixMicroC | 7 +++++++ 1 file changed, 7 insertions(+) create mode 100755 bin/hicBuildMatrixMicroC diff --git a/bin/hicBuildMatrixMicroC b/bin/hicBuildMatrixMicroC new file mode 100755 index 00000000..c648daa7 --- /dev/null +++ b/bin/hicBuildMatrixMicroC @@ -0,0 +1,7 @@ +#!/usr/bin/env python +# -*- coding: utf-8 -*- + +from hicexplorer.hicBuildMatrixMicroC import main + +if __name__ == "__main__": + main() From eeddf4201cb4fa42579f945588730700bb418083 Mon Sep 17 00:00:00 2001 From: Joachim Wolff Date: Wed, 27 Nov 2024 17:56:36 +0100 Subject: [PATCH 4/6] Fixing doc string test issue of different python versions --- hicexplorer/hicPlotDistVsCounts.py | 107 +++++++++++++++++++---------- 1 file changed, 72 insertions(+), 35 deletions(-) diff --git a/hicexplorer/hicPlotDistVsCounts.py b/hicexplorer/hicPlotDistVsCounts.py index 4a441641..1dca5ba0 100644 --- a/hicexplorer/hicPlotDistVsCounts.py +++ b/hicexplorer/hicPlotDistVsCounts.py @@ -5,6 +5,7 @@ import numpy as np import pandas as pd import argparse +import sys from hicmatrix import HiCMatrix from hicexplorer._version import __version__ @@ -138,41 +139,6 @@ def compute_distance_mean(hicmat, maxdepth=None, perchr=False, custom_cut_interv and values are ordered dictionary where keys are distances and values are a 2-tuple with the mean and the number of bins considered. - >>> from scipy.sparse import csr_matrix - >>> cut_intervals = [('a', 0, 10, 1), ('a', 10, 20, 1), - ... ('a', 20, 30, 1), ('a', 30, 40, 1), - ... ('b', 20, 30, 1), ('b', 30, 40, 1), ('b', 40, 50, 1)] - >>> hic = HiCMatrix.hiCMatrix() - >>> hic.nan_bins = [] - >>> matrix = np.array([ - ... [ 1, 8, 5, 3, 0, 0, 0], - ... [ 0, 4, 15, 5, 1, 0, 0], - ... [ 0, 0, 0, 7, 2, 0, 1], - ... [ 0, 0, 0, 0, 1, 0, 2], - ... [ 0, 0, 0, 0, 10, 0, 20], - ... [ 0, 0, 0, 0, 0, 0, 0], - ... [ 0, 0, 0, 0, 0, 0, 6]]) - - >>> hic.matrix = csr_matrix(matrix) - >>> hic.setMatrix(hic.matrix, cut_intervals) - >>> compute_distance_mean(hic) - {'all': OrderedDict({0: (3.0, 7), 10: (6.0, 5), 20: (10.0, 3), 30: (3.0, 1)})} - >>> compute_distance_mean(hic, perchr=True) - {'a': OrderedDict({0: (1.25, 4), 10: (10.0, 3), 20: (5.0, 2), 30: (3.0, 1)}), 'b': OrderedDict({0: (5.333333333333333, 3), 10: (0.0, 2), 20: (20.0, 1)})} - >>> custom_cut = [('tad1', 0, 10, 1), ('tad1', 10, 20, 1), ('tad2', 0, 10, 1), - ... ('tad2', 10, 20, 1), ('tad3', 0, 10, 1), ('tad3', 10, 20, 1), ('tad3', 20, 30, 1)] - >>> compute_distance_mean(hic, custom_cut_intervals=custom_cut) - {'all': OrderedDict({0: (3.0, 7), 10: (3.75, 4), 20: (20.0, 1)})} - >>> compute_distance_mean(hic, perchr=True, custom_cut_intervals=custom_cut) - {'a': OrderedDict({0: (1.25, 4), 10: (7.5, 2)}), 'b': OrderedDict({0: (5.333333333333333, 3), 10: (0.0, 2), 20: (20.0, 1)})} - >>> custom_cut = [('_ignore_0', 0, 10, 1), ('0', 0, 10, 1), - ... ('0', 10, 20, 1), ('_ignore_3', 0, 10, 1), - ... ('1', 0, 10, 1), ('1', 10, 20, 1), ('1', 20, 30, 1)] - >>> compute_distance_mean(hic, custom_cut_intervals=custom_cut) - {'all': OrderedDict({0: (4.0, 5), 10: (5.0, 3), 20: (20.0, 1)})} - - # >>> compute_distance_mean(hic, custom_cut_intervals=custom_cut, perchr=True) - # {'a': OrderedDict([(0, (2.0, 2)), (10, (15.0, 1))]), 'b': OrderedDict([(0, (5.333333333333333, 3)), (10, (0.0, 2)), (20, (20.0, 1))])} """ binsize = hicmat.getBinSize() @@ -515,3 +481,74 @@ def main(args=None): plt.tight_layout() plt.savefig(args.plotFile.name, bbox_inches='tight', bbox_extra_artists=(lgd,)) plt.close(fig) + + +if sys.version_info >= (3, 12): + compute_distance_mean.__doc__ += """ + >>> from scipy.sparse import csr_matrix + >>> cut_intervals = [('a', 0, 10, 1), ('a', 10, 20, 1), + ... ('a', 20, 30, 1), ('a', 30, 40, 1), + ... ('b', 20, 30, 1), ('b', 30, 40, 1), ('b', 40, 50, 1)] + >>> hic = HiCMatrix.hiCMatrix() + >>> hic.nan_bins = [] + >>> matrix = np.array([ + ... [ 1, 8, 5, 3, 0, 0, 0], + ... [ 0, 4, 15, 5, 1, 0, 0], + ... [ 0, 0, 0, 7, 2, 0, 1], + ... [ 0, 0, 0, 0, 1, 0, 2], + ... [ 0, 0, 0, 0, 10, 0, 20], + ... [ 0, 0, 0, 0, 0, 0, 0], + ... [ 0, 0, 0, 0, 0, 0, 6]]) + + >>> hic.matrix = csr_matrix(matrix) + >>> hic.setMatrix(hic.matrix, cut_intervals) + >>> compute_distance_mean(hic) + {'all': OrderedDict({0: (3.0, 7), 10: (6.0, 5), 20: (10.0, 3), 30: (3.0, 1)})} + >>> compute_distance_mean(hic, perchr=True) + {'a': OrderedDict({0: (1.25, 4), 10: (10.0, 3), 20: (5.0, 2), 30: (3.0, 1)}), 'b': OrderedDict({0: (5.333333333333333, 3), 10: (0.0, 2), 20: (20.0, 1)})} + >>> custom_cut = [('tad1', 0, 10, 1), ('tad1', 10, 20, 1), ('tad2', 0, 10, 1), + ... ('tad2', 10, 20, 1), ('tad3', 0, 10, 1), ('tad3', 10, 20, 1), ('tad3', 20, 30, 1)] + >>> compute_distance_mean(hic, custom_cut_intervals=custom_cut) + {'all': OrderedDict({0: (3.0, 7), 10: (3.75, 4), 20: (20.0, 1)})} + >>> compute_distance_mean(hic, perchr=True, custom_cut_intervals=custom_cut) + {'a': OrderedDict({0: (1.25, 4), 10: (7.5, 2)}), 'b': OrderedDict({0: (5.333333333333333, 3), 10: (0.0, 2), 20: (20.0, 1)})} + >>> custom_cut = [('_ignore_0', 0, 10, 1), ('0', 0, 10, 1), + ... ('0', 10, 20, 1), ('_ignore_3', 0, 10, 1), + ... ('1', 0, 10, 1), ('1', 10, 20, 1), ('1', 20, 30, 1)] + >>> compute_distance_mean(hic, custom_cut_intervals=custom_cut) + {'all': OrderedDict({0: (4.0, 5), 10: (5.0, 3), 20: (20.0, 1)})} + """ +else: + compute_distance_mean.__doc__ += """ + >>> from scipy.sparse import csr_matrix + >>> cut_intervals = [('a', 0, 10, 1), ('a', 10, 20, 1), + ... ('a', 20, 30, 1), ('a', 30, 40, 1), + ... ('b', 20, 30, 1), ('b', 30, 40, 1), ('b', 40, 50, 1)] + >>> hic = HiCMatrix.hiCMatrix() + >>> hic.nan_bins = [] + >>> matrix = np.array([ + ... [ 1, 8, 5, 3, 0, 0, 0], + ... [ 0, 4, 15, 5, 1, 0, 0], + ... [ 0, 0, 0, 7, 2, 0, 1], + ... [ 0, 0, 0, 0, 1, 0, 2], + ... [ 0, 0, 0, 0, 10, 0, 20], + ... [ 0, 0, 0, 0, 0, 0, 0], + ... [ 0, 0, 0, 0, 0, 0, 6]]) + >>> hic.matrix = csr_matrix(matrix) + >>> hic.setMatrix(hic.matrix, cut_intervals) + >>> compute_distance_mean(hic) + {'all': OrderedDict([(0, (3.0, 7)), (10, (6.0, 5)), (20, (10.0, 3)), (30, (3.0, 1))])} + >>> compute_distance_mean(hic, perchr=True) + {'a': OrderedDict([(0, (1.25, 4)), (10, (10.0, 3)), (20, (5.0, 2)), (30, (3.0, 1))]), 'b': OrderedDict([(0, (5.333333333333333, 3)), (10, (0.0, 2)), (20, (20.0, 1))])} + >>> custom_cut = [('tad1', 0, 10, 1), ('tad1', 10, 20, 1), ('tad2', 0, 10, 1), + ... ('tad2', 10, 20, 1), ('tad3', 0, 10, 1), ('tad3', 10, 20, 1), ('tad3', 20, 30, 1)] + >>> compute_distance_mean(hic, custom_cut_intervals=custom_cut) + {'all': OrderedDict([(0, (3.0, 7)), (10, (3.75, 4)), (20, (20.0, 1))])} + >>> compute_distance_mean(hic, perchr=True, custom_cut_intervals=custom_cut) + {'a': OrderedDict([(0, (1.25, 4)), (10, (7.5, 2))]), 'b': OrderedDict([(0, (5.333333333333333, 3)), (10, (0.0, 2)), (20, (20.0, 1))])} + >>> custom_cut = [('_ignore_0', 0, 10, 1), ('0', 0, 10, 1), + ... ('0', 10, 20, 1), ('_ignore_3', 0, 10, 1), + ... ('1', 0, 10, 1), ('1', 10, 20, 1), ('1', 20, 30, 1)] + >>> compute_distance_mean(hic, custom_cut_intervals=custom_cut) + {'all': OrderedDict([(0, (4.0, 5)), (10, (5.0, 3)), (20, (20.0, 1))])} + """ From d7bb826c284139a3ceeb934519aaefb276bf1ea1 Mon Sep 17 00:00:00 2001 From: Joachim Wolff Date: Wed, 27 Nov 2024 19:20:35 +0100 Subject: [PATCH 5/6] Try to fix azure with macOS --- requirements.txt | 1 + 1 file changed, 1 insertion(+) diff --git a/requirements.txt b/requirements.txt index 4e8e3511..2f83dc0f 100644 --- a/requirements.txt +++ b/requirements.txt @@ -26,3 +26,4 @@ python-graphviz >= 0.20 scikit-learn >= 1.3,<1.4 imbalanced-learn >= 0.11 cleanlab >= 2.5 +xz \ No newline at end of file From 1238d17cd17ee7521dc811704a7829682fbc9564 Mon Sep 17 00:00:00 2001 From: Joachim Wolff Date: Wed, 27 Nov 2024 19:33:09 +0100 Subject: [PATCH 6/6] Update docs, remove of macOS fix try --- README.rst | 6 +++--- docs/content/list-of-tools.rst | 3 +++ requirements.txt | 3 +-- 3 files changed, 7 insertions(+), 5 deletions(-) diff --git a/README.rst b/README.rst index e2f1ed3f..69b0cf9c 100644 --- a/README.rst +++ b/README.rst @@ -16,8 +16,8 @@ HiCExplorer =========== -Set of programs to process, analyze and visualize Hi-C and cHi-C data ---------------------------------------------------------------------- +Set of programs to process, analyze and visualize Hi-C, Micro-C and cHi-C data +------------------------------------------------------------------------------ Sequencing techniques that probe the 3D organization of the genome generate large amounts of data whose processing, analysis and visualization is challenging. Here, we present HiCExplorer, a set of tools for the analysis and @@ -26,7 +26,7 @@ of contacts, TAD detection, A/B compartments, merging, reordering or chromosomes `cooler `_ and detection of long-range contacts. Moreover, it allows the visualization of multiple contact matrices along with other types of data like genes, compartments, ChIP-seq coverage tracks (and in general any type of genomic scores), long range contacts and the visualization of viewpoints. - +With version 3.7.6 we introduce the support for Micro-C data for the build of matrices. Single-cell Hi-C data --------------------- diff --git a/docs/content/list-of-tools.rst b/docs/content/list-of-tools.rst index 6cd92433..12b4347f 100644 --- a/docs/content/list-of-tools.rst +++ b/docs/content/list-of-tools.rst @@ -8,6 +8,7 @@ Tools for Hi-C data pre-processing tools/hicFindRestSite tools/hicBuildMatrix + tools/hicBuildMatrixMicroC tools/hicSumMatrices tools/hicMergeMatrixBins tools/hicCorrectMatrix @@ -98,6 +99,8 @@ For single-cell Hi-C data analysis please use `scHiCExplorer = 0.2.7 python-graphviz >= 0.20 scikit-learn >= 1.3,<1.4 imbalanced-learn >= 0.11 -cleanlab >= 2.5 -xz \ No newline at end of file +cleanlab >= 2.5 \ No newline at end of file