Introduction of Interlab
Reliable and repeatable measurement plays an important role as an engineering discipline in synthetic biology. The InterLab Study aims to identify and correct the sources of systematic variability in synthetic biology measurement, thus to improve the measurement tools and methods which can enable labs to reliably build upon others’ work. It will promote the collaboration within the synthetic biology community. Hence, it may propel the synthetic biology to achieve its full potential.
The previous InterLab studies showed that measuring GFP expression in absolute fluorescence units calibrated against a known concentration of fluorescent molecule can greatly reduce the variability in different labs’ measurements.
The Fifth Year InterLab’s goal is to remove another source of variability, the number of cells in the sample, by using new cell-counting method.
GreatBay_China investigation had helped in answering the question:Can we reduce lab-to-lab variability in fluorescence measurements by normalizing to absolute cell count or colony-forming units (CFUs) instead of OD?
Supplies and Reagents
DNA/Plasmids
- Negative Control: BBa_R0040
- Positive Control: BBa_I20270
- Test Device 1: BBa_J364000
- Test Device 2: BBa_J364001
- Test Device 3: BBa_J364002
- Test Device 4: BBa_J364007
- Test Device 5: BBa_J364008
- Test Device 6: BBa_J364009
Apparatus
- 96 well plates, black with clear flat bottom (provided by Bluepha Lab)
- Plate reader
- Flow Cytometry
- Incubator at 37˚C
- Ice bucket with ice
- Foil covered 50 ml tube
- Eppendorf tubes
- Micropipettes
- Micropipette tips
Materials
Measurement Kit
- LUDOX CL-X
- Silica beads
- Fluorescein
1x PBS (Phosphate buffered saline, pH 7.4-7.6)
ddH2O
Competent cells (Escherichia coli strain DH5α)
LB (Luria Bertani) media
Chloramphenicol
LB plates
distilled water
