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Gel Electrophoresis and PCR Round 2
This week we were able to run our PCR samples from 9/27 on a 1% agarose gel. All of our samples turned out very well - our DNA samples amplified around 1,000 bp (we were expecting 1,044 bp) and our controls were clean. There was no visible banding patterns within our negative control and the positive control displayed a strong band around 400 bp.
Well 12 is the DNA 100bp Plus marker, wells 13-15 are pure A DNA samples, wells 16-18 are pure B DNA samples, 19 is our negative control, and 20 is our positive control. Samples A & B are from the same organismal source but two different extractions. Wells 13 & 16 had annealing temperatures of 65°C. Wells 14 & 17 had 59.1°C and Wells 15 & 18 had 55.5°C. This was to determine the best temperature for our extended primers.
Thursday, 10/4, we ran 10 more PCR samples in order to have a larger quantity of available DNA to purify. We chose the sample set from Well 13 (Pure A sample, annealing temp 65°C) because it came out the strongest on the gel. Along with the 10 samples, we are again including a positive and negative control which are exactly the same as our first PCR controls. We are also using the same thermocycler protocol as before.