British_Columbia_Base

The purpose of iGEM’s Interlab study is to establish a standardized GFP measurement protocol that is universal and can produce the same results despite using different plate readers. In this year’s Interlab study, RBS devices (BCDs) were also measured by various iGEM teams. Teams that participate in the Interlab study this year are required to follow a standardized procedure and submit their data using a plate reader.

The official protocol for the Interlab study can be found on the official iGEM Interlab Study page here. Our data for the Interlab Study is as follows:

Methods

Transformations

The eight plasmids (Positive Control, Negative Control, Test Device 1, Test Device 2, Test Device 3, Test Device 4, Test Device 5, and Test Device 6) were transformed into competent E.coli DH5-alpha cells using the E.coli Chemical Transformation protocol here. The transformants were then plated on LB+25 mg/mL Chloramphenicol plates and incubated overnight at 37 degrees Celsius. Two colonies were picked from each plate and inoculated in 5 mL of LB + 25 mg/mL Chloramphenicol overnight at 37 degrees Celsius and 220 rpm.

Calibration

The OD600 reference point and fluorescein standard curves were prepared following the official Interlab Study protocol, which can be found here.
This was what our standard fluorescence curves looked like:

Measurements

The next day after the inoculation, we measured the OD600 of the overnight cultures using a spectrophotometer. The cultures were diluted to a target OD600 of 0.02 using the Dilution Calculation Sheet, which can be found here. We then incubated the cultures at 37 degrees Celsius and 220 rpm. We took measurements of the cultures at 0, 2, 4, and 6 hours of incubation by pipetting 500 uL of each culture into wells on a 96-well plate according to the assigned layout in the protocol. We made a modification to the protocol by measuring the OD600 and fluorescence after each time interval, instead of measuring all of the time points at the very end.

Results

Detailed Results

The detailed results of our Interlab Study can be found here.