Team:iTesla-SoundBio/Results

Results

(Left) The gel shows a comparison of our digested RFP (wells 3-4 from the left) to our undigested RFP (well 8).
(Right) An example showing how well our PCRs worked. (From the top down) Well 3 is pcbA5 (biobrick insert), Well 5 is pcbA5 (pet29b insert), Well 7 (faint band) is our promoter, and Well 9 (very faint band) is pSB1C3.



We ultimately sent in two different biobricks: the upstream region of genes pcbA4/5 (BBa_K2363001) and the pcbA5 gene itself (BBa_K2363000). Our control was the digested backbone vector without insert added in the ligation reaction. With 100% efficiency, we would not expect any colonies to grow on the control plate, but the level of background we observed was minimal. The BBa_K2363001 transformation plate had significant number of white colonies, suggesting the ligations and transformations were effective. We chose to miniprep and sequence two of the white colonies to make sure they were not false positives. There were a few red colonies on the plate which was consistent with the background observed in our control plate. Our pcbA5 biobrick didn’t show as many colonies, and there were more red colonies than white colonies. Nevertheless, we went on to miniprep and sequence two of the white colonies. Sequencing revealed that both the promoter and pcbA5 biobricks obtained from our selected colonies perfectly matched our expected constructs. No point mutations were observed. See raw sequencing data attached here for BBa_K2363001 and here for BBa_K2363000

Control


BBa_K2363001


BBa_K2363000