This week, I tested a set of primers which I designed last spring but had not yet used in a PCR. These primers, which I call P-CAT-3f and P-CAT-3r, were created during the same sessions that produced the P-CAT-4 primer set which I have been using to amplify DNA this summer.
As you may recall, the primer set sequences I am using were identified after I analyzed the 16s ribosomal gene maps of three gram negative bacterial strains which we have here in the lab. I then began this phase of my project by testing one primer set against nine gram negative species, including the original three used to design the primers. When several PCRs were run and DNA was amplified, I then tested the primers against six gram positive species, which also resulted in DNA amplification.
For this week's test, I first performed an extraction on the six gram positives I am using, then ran a PCR using the aforementioned P-CAT-4 set. After running a gel and verifying that DNA was present, I then took the original extraction sample and set up a new PCR using the P-CAT-3 primer set.
The results? Five of the six species amplified. The sixth, E. faecalis, did not (see gels below). Base pair sizes were approximated at 350-425 BPs long.
That's it for this week. Until next week, please enjoy this lecture from Dr. Jennifer Doudna, who co-discovered the CRISPR gene editing technique. Cheers!
https://www.youtube.com/watch?v=SuAxDVBt7kQ
Gel results:
No comments:
Post a Comment