Team:Nottingham/InterLab
InterLab
The aim of the iGEM InterLab study is to work towards a more reliable and repeatable measurement system to make synthetic biology an engineering biology. All participating laboratories first calibrated their instruments by obtaining standard curves using sodium fluorescein which was provided in our kits. This allowed us to fix settings such as top/bottom optic, gain and type of plate used so that all conditions were the same for our GFP and CFU protocols.
Table 1 shows data for the OD600 reference point.
Table 2 shows data for the particle standard curve (standard curve below).
Table 3 shows data for fluorescein standard curve (standard curve below).
(Blue cells were raw data measurements, gold cells were calculated) After the machine was calibrated fluorescence and absorbance of GFP in 8 different devices was measured using the standardised method provided by iGEM. The results gathered were suggestive of which device had the highest strength of gene expression. The data collection sheet provided converted our raw plate reader measurements into arbitrary fluorescein and abs600 values as well as calculating the µM fluorescein per OD as shown in the following tables.
Tables 4 and 5 show calculated values of µM fluorescein per OD at 0 hour and 6 hour time points. This was calculated from OD measurements we took before beginning the assay.
Tables 6 and 7 show calculated arbitrary values of net fluorescein at 0 hour and 6 hour time points.
Tables 8 and 9 show net values of absorbance of light (600nm) at 0 hour and 6 hour time points.
What did we gain from our InterLab experience? The InterLab study has allowed us to contribute to making synthetic biology more accurate through providing a series of standardised procedures of calibration and measurement tests. It helped our project as we were then able to use our new calibrated programmes to compare our promoters GFP assay data with iGEM standard data.
