This week, I continued repetitive testing of my P-CAT-4 primer set which had shown promise in some earlier test rounds. This was in order to verify that I am receiving the same positive PCR amplification results from separate test runs.
I performed another simple heat extraction on the gram positives (and a Pseudomonas aueruginosa strain that had been causing me some difficulty in a previous round) that I am using for this study, thawed out my previously tested gram negatives, and set up a final PCR (of the session) using that primer set. I then ran a standard electrophoresis gel @100 V for 30 minutes.
The results? Inconclusive. P. aeruginosa showed post-PCR gel banding, but there was a variation in gram-negative results from all previously performed PCRS using the samples I had backstocked. Due to these results, I ordered more stock of these gram-negative species and intend to re-test them for DNA amplification when the next semester begins as I am concerned that the samples may have been corrupted by the length of time they have been in storage.
I simply ran out of time during this session to complete this portion of the study.
Here is a P. aeruginosa gel with a ladder from this week's session; the referenced sample is all the way to the right.
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