840:153g:Projects/project26/2012/11/01

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Cloning of Atrolysin A Main Project Page
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DNA Extraction

  • On Tuesday of this week we ran a PCR of our DNA sample. Thursday we checked it on a gel and there was a high concentration down on the lanes. We then realized we used the Stock Solution of our primers instead of our Working Solution. We then set up another PCR reaction using the Working Solution of primers. Then next Tuesday we will check it on a gel. We have a couple of things to try. One is to put our gene into a T-vector and get sequenced. The other is if the PCR works then we will try a PCR with the extended primers and try and put our gene into the PSB vector. This all depends on what we find on Tuesday.

Image:11-1-12A.tif

Starting on the left. Lanes are: A1, E1, A2, E2, A3, Ladder, E3, Negative control, Positive control. The "A" samples contain 5 microliters of DNA and the "E" samples contain 0.5 microliters of DNA. The "1" samples were placed at 65 degrees C in the PCR, the "2" samples were placed at 55 degrees C, and the "3" samples and the negative and positive controls were placed at 50 degrees C. There were no bands present which means that the amplification did not work.

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