Team:ShanghaiTech/InterLab
Introduction
Reliable and reproducible measurement is a key component to all engineering disciplines. The same holds true for synthetic biology. However, it is difficult to repeat the measurement data from different labs. The goal of the iGEM InterLab Study is to identify and correct the sources of systematic variability in synthetic biology measurements, so eventually, measurements that are taken in different labs will be no more variable than measurements taken within the same lab.
This year, the objective of the InterLab study is to test if we can reduce lab-to-lab variability in fluorescence measurements by normalizing to absolute cell count or colony-forming units (CFUs) instead of OD reading. Teams from around the world are using standardized biological parts, bacterial strain and measurement procedures provided in a detailed protocol by iGEM.
Results and Discussion
Calibration
Fig.1 OD600 reference point
Fig.2 Particle standard curve
Fig.3 Fluorescein standard curve
Cell measurement
All the raw data and following tables refer to the well arrangement of iGEM protocal below.
Fig.4 Well arrangement
Fig.5 Abs600 Raw Readings
After 18 hours growth at 37°C and 220 rpm orbital rotation speed, cultures were diluted to a target Abs600 of 0.02. After deleting background, 0h time point data has an average Abs600 of 0.0206. The rest of the diluted cultures were incubated at 37°C and 220 rpm for 6 hours, and after 6 hours of growth, significant Abs600 increase could be detected.
Fig.6 Fluorescence Raw Readings
At 0h time point no fluorescence or trace fluorescence were detected. At 6h time point, negative control still remain low fluorescence, though the fluorescence in Device 3 and Device 5 behaved unexpected low, yet all other 6 groups appeared apparent increase of fluorescence.
Fig.7 CFUs per 0.1 OD600 E. coli cultures
Extra Credit
Fig.8 Flow Cytometry
After measuring each plate with the plate reader, we also collected data from all wells by using flow cytometer. At least 10,000 events per well were collected.
Instrument information: CytoFlex S
Full data link: link