Week 12-Limited results

Today, I ran an electrophoresis gel on the six PCR samples that I created last week. Once again, I was reminded that a hurried project is a failed project, as I had to load three gels due to a calculation error in my sample sizes. The error occurred as a result of me attempting to rush through the loading process, instead of taking the time to check and then re-check my samples.

While the PCR process worked, it was not at optimal amplification and my gel was muddled. The likely culprit for this was probably the annealing temperature, which was set for all six samples at 43°C. The melting temperatures for my primers ranged from 48°C to 60°C. I had set the temperature to accommodate the lowest melting temperature of my four primers, in order to run all six samples through the thermal cycler during one PCR. Next week, I will repeat the PCR, but at that time I will run separate cycles depending on the primer samples, so that I can more closely align the annealing temperatures with the utilized primers.

Here is an image of the gel results: