Team:Tuebingen/Measurement
InterLab Study
Fluorescence measurement is a method abundant in all major labs.
Following up on the past years’ success, the iGEM HQ again challenged teams to take part in the InterLab study.
Measuring the fluorescence of various gene constructs becomes more and more important during everyday lab work. However, the comparability in different labs is still under debate. Using the fact that teams all over the world participate in iGEM, this offers the perfect opportunity to see how results of the same experiment performed all over the world compare. This year’s aim was to compare the fluorescence yield of five different constructs containing GFP under control of different RBS and promoters. By using a FITC standard curve and LUDOX to correct for errors in OD600 measurements, the results of all teams can be easily compared.
Materials and Methods
The following five constructs from the InterLab kit were used to transform E. coli DH10beta:
FITC and LUDOX standards were measured in a TECAN plate reader, using 96-well plates with transparent bottoms. LUDOX was measured at 600nm with a bandwidth of 9 nm and 25 flashes. FITC was excited at 488 nm (bandwidth 9 nm) and emission was recorded at 520 nm (bandwidth 20nm) with a fixed gain of 50 and 25 flashes. Identical settings were used for measurement of the devices.
All measurements were done as described in the Interlab plate reader protocol (see Methods). Transformations were performed as described here.
