Part:BBa_K3773520:Experience

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iGEM21_William_and_Mary

Our initial confirmation of this part's function was as follows. To test the functionality of our circuits, we inoculated from our -20ºC glycerol stocks and let the cultures grow overnight. We then pipetted three wells, each containing 200 µL of our culture, into a 96-well plate, along with three wells of the same volume of each of our positive and negative controls. Our positive control was WM21_013, a circuit that we had already found to exhibit incredibly high fluorescence values, and our negative control was LB, as the medium we used should not have been fluorescent. We then ran the plate through a Synergy H-1 plate reader, and determined whether the wells containing our plasmid cultures were more fluorescent than the negative control, with the positive control as a benchmark for the expected high level of fluorescence given our use of sfGFP. Unfortunately, both of the secretion circuits (K3773519 and K3773520) failed to be functionally confirmed.

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