![]( https://static.igem.org/mediawiki/2017/a/af/InterLB.png)
Introduction The Fourth International InterLaboratory Measurement Study seeks to establish a reproducible plate reader-based GFP measurement protocol. Eight plasmids were transformed into competent DH5a E.coli cells and expressed for varying lengths of time. The optical density and fluorescence of each device was recorded over a 6 hour period. The major goal of this collective effort is to answer the following question: how close can the numbers be when fluorescence is measured all around the world?
Materials and Methods In this Interlab study, the following 8 RBS devices were provided in the Kit Plate 7 in the 2017 Distribution Kit:
- Positive control
- Negative control
- Test Device 1:J23101.BCD2.E0040.B0015
- PTest Device 2: J23106.BCD2.E0040.B0015
- Test Device 3: J23117.BCD2.E0040.B0015
- Test Device 4: J23101+I13504
- Test Device 5: J23106+I13504
- Test Device 6: J23117+I13504
- 2 uL DNA were added to 50 uL competent DH5a
- The incubation times on ice before and after the heat shock were 20 minutes and 2 minutes, respectively.
- The heat shock was performed for 90 seconds instead of 50 seconds.
- 800 uL SOC broth were added to each transformation.
- After the one-hour shaking incubation, each transformation tube was centrifuged for 1 minute, 13,000 rpm. Afterwards, 700 uL supernatant were discarded and the pellet was resuspended with the remaining 100 uL before being plated onto LB agar plate.
Results The results of the Interlab Study were tabulated in the following link.