PCR Results from 7/10
Here are the results from the 24 PCR reactions we did yesterday. About 2/3 of the reactions used PCC 7942 DNA and Peng's primers for amplifying the KaiABC region. The rest were positive controls using BioBrick inserts and primers.
It appears that the 2 mM Mg+ buffer produced the best results. We're not sure whether an annealing temperature of 55° or 65° produces better yield, since our 65° machine was accidentally misprogrammed.
Reactions 17 and 21 yielded bands in the 3kb region, which is the length of our KaiABC extract.
We extracted and purified the 3kb band from reactions 21. According to the nanodrop, we had a DNA concentration of 9.5 ng / µL.
Second gel electrophoresis
We ran a second gel electrophoresis with reactions 17 and 21 as backups in case our transformation failed (see next entry). The results are shown below. We extracted reaction 21 (third lane) and refrigerated it (it hasn't been purified yet).