Made another 1% gel (0.5 grams + 50mL TAE buffer) in the Fisher Lab electrophoresis rig. Used the teaching lab 10-well comb because the Fisher Lab doesn't own any combs of that size.
Loaded samples from 7-9-14 that were amplified using the AFB primer sets. (2 uL EZ Vision + 15 uL sample, loaded 7 uL into each well). Ran the gel at 120V for 30 minutes. Upon imaging, there were absolutely no bands that appeared on the gel and even the DNA marker wasn't very apparent. For some reason I had nearly a blank gel.
The amplicons had been stored at -20C for 4 days. I am confident that the samples were loaded successfully into the wells, that they were thoroughly mixed with the loading dye, and that they ran the correct way across the gel. I am not sure what caused the previously visible amplicons to no longer appear on the gel. My only conclusion is to repeat the amplification and try again (soon!).
In the future, I will re-PCR amplify each of the samples using the AFB primers and visualize via gel electrrophoresis. DNA lysates from each sample have been stored at -20C.
//EWW
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