Using the three color calibration protocol: Does the order of transcritional units influence their expression strength?
In this experiment, your team will measure the fluorescence of six devices that encode two fluorescence proteins in two transcriptional units. The devices differ in the order of the transcriptional units. You will calibrate the fluorescence of these devices to the calibrant dyes and the optical density of the culture to the cell density calibrant.
This experiment aims to assess the lab-to-lab reproducibility of the three color calibration protocol when two fluorescent proteins are expressed in the same cell. Besides this technical question, it also adresses a fundamental synthetic biology question: does the order of the transcritional units (that encode for the two different fluorescent proteins) on the devices influence their expression levels?
- E. coli DH5 alpha
- LB Broth+chloramphenicol
- chloramphenicol
- Negative control 2022
- Positive control 2022 Green
- Positive control red (mCherry) Exp 2
- Test Device 1 Exp 2 (Dual construct Green and Blue)
- Test Device 2 Exp 2 (Dual construct Green and Red)
- Test Device 3 Exp 2 (Dual construct Red and Blue)
- Test Device 4 Exp 2 (Dual construct Blue and Red)
- Test Device 5 Exp 2 (Dual construct Red and Green)
- culture tube (x 32)
- 1.5 mL microfuge tube (x 48)
- 50 ml conical tube (x 16)
- 96 well plate (x 2)
| Part | Coordinate |
|---|---|
| BBa_J428100 | Kit Plate 1 Well 12M |
| 3_Colors_ins_K2656022 | BBa_J428112 Kit Plate 1 Well 14C |
| BBa_J428101 | Kit Plate 1 Well 12I |
| BBa_J428106 | Kit Plate 1 Well 12G |
| BBa_J428107 | Kit Plate 1 Well 3L |
| BBa_J428105 | Kit Plate 1 Well 5J |
| BBa_J428108 | Kit Plate 1 Well 14E |
| DC_R_ins_K2656022 | BBa_J428104 Kit Plate 1 Well 5L |
- Import data into the provided Excel file: .
Timestamp: 2023-03-11 16:05:34.526107 Protocol version: 1.0b