For the 2018 Interlab, it mainly focuses on reducing the lab-to-lab variability in fluorescence measurements by normalizing to colony-forming units (CFUs) instead of OD. Our team participated in this study for creating a standardize condition for all laboratory environments.
We transformed Escherichia coli DH5α with the plasmids that are listed below and were incubated at 37 C for 16 hours.
Before the cell measurement protocol, there are three sets of calibration measurements be taken. We did an OD600reference point that enable us to obtain a conversion factor to transform absorbance(Abs600) data from plate reader into a comparable OD600 measurement for the spectrophotometer. A particle standard curve, help to convert Abs600 measurements to an estimated number of cells. In addition, a fluorescein standard curve that is necessary when comparing fluorescence output of test devices between teams.
We measure both Abs600and fluorescence for the two colonies, with one colony at 0 hour and another at 6 hours.
We used the given formula to count colonies for two Positive Control (BBa_I20270) cultures and two Negative Control (BBa_R0040) cultures.
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