This year, the BrockU iGEM team successfully participated in the fifth annual InterLab study. For this project, we determined the fluorescence of E. coli cells carrying different plasmids containing GFP (Figure 1). We also determined the colony-forming units of each of these bacterial cultures (Figure 2). This was done as a part of a larger study to identify variability in synthetic biology measurements, and attempt to correct this variability.
The BrockU iGEM team saw the InterLab study as a great way compare our measurement techniques against labs around the world! We also saw the value in attempting to standardize measurement techniques globally, as this would allow for protocols to be easily replicated in any laboratory in the world.
The BrockU iGEM team saw the InterLab study as a great way compare our measurement techniques against labs around the world! We also saw the value in attempting to standardize measurement techniques globally, as this would allow for protocols to be easily replicated in any laboratory in the world.
Figure 1: Fluorescence per Particle of Various Test Devices. As per the InterLab protocol, we transfected E. coli with various test devices containing GFP, as well as a positive and negative control. The cells were cultured overnight, and the GFP signal was recorded at two time points, time 0h and time 6h; alongside standards for measuring fluorescence and cell size. Using InterLab's protocol, we determined the fluorescence per particle in each of the test device cultures. Although these results were not quite what we expected, we observed some slight variation in fluorescence per particle across the different test devices.
Figure 2: Colony Forming Units Assay. In order to determine the colony forming units of the transfected E. coli cultures, serial dilutions were performed on samples of the positive and negative controls from a standard OD. By counting the number of colonies on each of the diluted plates, we were able to calculate the amount of colony forming units of the bacteria.