From OpenWetWare
Jump to: navigation, search

Today we transformed our plasmids (3 different ones containing the 3 different promoter parts and our gene) into competent E. coli cells. After the transformation procedure, we plated each transformation sample, a positive control (cells transformed with a standard plasmid), a negative control (cells with PCR water only), and pure competent cells onto amp plates. Onto sections of a regular LB plate, we plated our negative control and the pure competent cells, because these were the only samples that should be unable to grow on the amp plates, and we needed a way to make sure the cells are alive. Next time we will be checking the growth on the plates to see if we have white colonies on the amp plates which would indicate successful transformation. We would then have to check that it was our plasmid that was taken up, however, by isolating the plasmid from the bacteria, running an RE digest, and running a gel to check for the correct band sizes. If we do not get successful growth on our plates, we will redo the ligation and transformation procedures.