This week, I have been repeating experiments I have previously done several times in order to either eliminate or account for an anomaly in my last round of gram-negative PCR amplifications. In initial testing, the primers I designed had amplified all 15 of the species chosen for my study. Multiple rounds of testing had positive post-PCR presence of DNA in my gels, but in a later phase of testing two of the gram-negatives did not amplify during PCR.
I am not certain at all why this occurred; there are a myriad of reasons which could account for the anomaly, including human error. So I did a new round of DNA extractions this week, ran some gels, and verified if DNA was present. All of the tested species were positive for DNA.
I subsequently banked the samples until next week, when I will be able to get some time on the thermocycler. Results of the PCR to follow on next week's blog.
Thanks for checking in with me.
Photo: Gel samples of 9 gram negative bacterial species showing DNA presence.
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