Part:BBa_J428112:Experience

Characterization of BBa_J428112

Lambert iGEM transformed BBa_J428112 into two different strains of E. coli (BL21 and DH5-alpha) to characterize the differences in fluorescence. BL21 is used for protein expression, whereas DH5-alpha is utilized for plasmid transformation. Therefore, we expect the test device transformed into the BL21 cells to produce more fluorescence than the test device transformed into the DH5-alpha cells. We characterized fluorescence expression of BBa_J428112 in BL21 and DH5-alpha using the iGEM InterLab Experiment 1 Protocol. The InterLab study instructs to quantify the sample at a 0 hour and 6 hour time mark. To better align with members’ schedules, we only quantified fluorescence at a 0 hour time mark.

As expected, BL21 exhibited statistically greater amounts of fluorescence than DH5-alpha (see Fig. 1). To evaluate the accuracy of our hardware component, we quantified the same samples in both plate reader and Micro-Q (see MicroQ). The values gathered from Micro-Q were scaled up by a factor of 104 to be comparable with those produced by the plate reader. The fluorescence/OD600 values are consistent between MicroQ and plate reader at 6766.657 and 7405.837 for DH5-alpha, and 16916.56 and 21328.17 for BL21. When comparing fluorescence in each cell strain individually, the error bars for MicroQ and plate reader overlap (see Fig. 1). Therefore, the outputs of MicroQ and plate reader are not significantly different, validating the fluorescence measurement of our hardware device.

Figure 1. Graph depicting characterization of BBa_J428112 in DH5-alpha and BL21 quantified in both MicroQ and Plate Reader. The error bars between DH5-alpha and BL21 do not overlap, suggesting there is a statistically significant difference. The error bars between MicroQ and Plate Reader do overlap, suggesting there is not a statistically significant difference.

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