Team:BGIC-Global/InterLab
Aim
Many scientists have tried to use relative expression comparisons to solve the incomparable fluorescence data problem. However, it is harder to debug engineered biological constructs and interpret your experimental controls when it is unable to directly compare measurements. Thus Interlab study provide researchers with a detailed protocol and data analysis form that yields absolute units for measurements of GFP in a plate reader to address these issues.
Materials and methods
The parts for the constructs used in the Interlab study (BBa_R0040, BBa_J20270, BBa_J364000, BBa_J364001, BBa_J364002, BBa_364007, BBa_364008 and BBa_364009) were taken from the iGEM distribution plates and transformed into E. coli DH5α strain and then measured the absorbance and fluorescence using Tecan Infinite M1000 plate reader. Inoculation and cultivation was done following the InterLab Protocol.
Results
As stipulated by the protocol, first we used the LUDOX and FITC provided to get standard measurements for both OD600 and Fluorescence. LODOX absorbance values are similar to example data and particle and fluorescence dilution create linear standard curve.
Device measurements
The fluorescence data of each constructs is tested at the time point of 0h and 6h following the protocol of Interlab. The colony forming units test is carried out according to the protocol as well.
Discussion
Colony 1 and 2 have similar Abs600 and fluorescence values which means that different colonies show the same phenotypes. The Device 4 and 6 show poorer performance than other devices. With the observation of our raw data, we can conclude that Device 1 and 4 show stronger fluorescence compared to the others while Device 3 and 5 show similar fluorescence strength as the negative control. The CFU of each dilution of the devices have the same magnitude as the example which means that the cell numbers of the overnight culture are similar.