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Parameters_template.txt
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This parameters file will guide the PrimedRPA-based primer and probe design process. Please follow the instructions outlined below:
----####----
Important Note - Do not remove any of the “>” and write your input directly after this symbol.
----####----
Please define the reference name for this PrimedRPA run:
>{}
Please indicate if you would like to use a previously generated Alignment File: [NO or File path]
>{}
Please indicate if you would like to use the previously generated Binding Sites: [NO or File path]
>{}
Please enter the path, from your current working directory, to the input fasta file:
>{}
Please classify the contents of the input fasta file as one of the following options: [SS, MS, AMS]. Whereby:
SS = Single sequence
MS = Multiple unaligned sequences
MAS = Multiple aligned sequences
>{}
If multiple sequences are present in the input fasta file (Classification of MS or MAS), please indicate below the
percentage identity required for the primers and probes target binding sites:
>{}
Please indicate if a primer identity anchor is required. [NO or length of anchor]
>{}
Desired primer length (This can be a range: 28-32 or fixed value: 32):
>{}
Please state if you require a probe to be designed and if so what type [NO,EXO,NFO]
>{}
Desired probe length (This can be a range: 45-50 or fixed value: 50):
>{}
Below please define your max amplicon length.
>{}
Below please state the repeat nucleotide cut-off in bp (e.g. 5bp will exclude sequences containing GGGGG).
>{}
Below please insert the minimum percentage GC content for primer/probe:
>{}
Below please insert the maximum percentage GC content for primer/probe:
>{}
Below please indicate the percentage match tolerance for primer-probe dimerisation and secondary structure formation:
>{}
Please enter [No or Path to Background file] below to identify if you want to perform a background DNA binding check:
>{}
Below please insert the percentage background cross reactivity threshold:
>{}
Below please indicate if you would like to implement a Background Hard Fail Filter [NO,YES]:
>{}
Please define the maximum number of sets you would like to identify:
>{}
Please define the number of threads available:
>{}
Blastn Cross Reactivity Search Settings [Basic or Advanced or Fast]
>{}
Blastn Evalue
>{}