User:Anthony Salvagno/Notebook/Research/2009/10/29/Redoing Unzipping: Anchor PCR of working reaction

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Note: I found some pRL574 1.5ng/ul in one of my boxes from Osley lab. And I found the pRL574 (not diluted) that I had saved for transformation. There may be like 5ul of that left. The pRL574 from Osley lab might not be so old, unless it comes from the same stock, but I don't think so.

Gel to come tomorrow. (Steve Koch 00:28, 30 October 2009 (EDT): Good luck!)


  1. 1x Ladder
  2. 2x Ladder
  3. 3x Ladder
  4. Rxn 1 - 12ul of PCR sample
  5. Rxn 2 - 12ul of PCR sample
  6. Rxn 3 - 12ul of PCR sample
  7. Rxn 4 - 12ul of PCR sample

On the left is the intensity profiles of the ladder (lanes 1-3) at the 3kb band, and the right is the intensity profiles of 3 of the PCR reactions (lanes 5-7). The PCR samples are more luminous than lane 3 (3x ladder), but I will just say it is equal to underestimate results. So in the 3kb band, there are 125ng of DNA. In my samples there are 12ul of PCR. 3 * 125/12=41.666ng/ul of PCR. I have 88 left for each so about 3.6ug of DNA in each tube. I have 5 tubes total (including the one from yesterday). I can do about 2 columns of PCR cleanup.

(Steve Koch 12:02, 30 October 2009 (EDT):YES! Did you use "block" or "sim tube?" Ant: I used "sim tube")