Part:BBa_K3773511:Experience

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iGEM21_William_and_Mary

Our initial confirmation of this part's function was as follows. After transformations, we attempted to confirm the functionality of our circuits. We did this by growing a culture of transformed DH5-alpha cells overnight (37°C and 250 RPM), diluting to an OD600 of 0.4, and following 2-4 hours of growth, incubating the cells with the aptamer-activating dye DFHBI-1T at room temperature for 45 minutes. After the 45-minute incubation period, we tested for aptamer fluorescence in our culture using a plate reader and comparing its fluorescence to samples of untransformed cells and LB (negative controls) and samples of our translational burden sensor circuit (positive control). However, we were unable to confirm the functionality of our circuits, as the fluorescence levels of our samples transformed with transcriptional burden sensor circuits was similar to that of the negative controls. This is most likely due to synthesis difficulties, as F30-Broccoli itself contains repeats within its sequence as well as a hairpin within its sequence.

User Reviews

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