User:Anthony Salvagno/Notebook/Research/2011/01/06/1.1kb Stretch PCR and Results
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I actually did the PCR yesterday but was a bad boy. I didn't think it was worth mentioning at all, but I'm turning over a new leaf. I'm going to start properly documenting my materials. I have to somehow link this PCR reaction which will be in some tube to this page, QR code?
PCR Reaction
{{#widget:Google Spreadsheet |key=0Agbdciapt4QZdEk5OWNUS2lxUFF6VkNsVmdzeUtDRUE |width=500 |height=300 }} This is my basic PCR template, but I used it literally for this reaction, using primers F834-dig (10/28/09) and R2008-biotin (11/19/10) and pRL574 for my template. I also used Platinum Taq from Invitrogen but kept everything else the same and used all the same final concentrations and volumes as the table indicates.
Oh yea, the program I used has a cycle of:
- 94C - 30s
- 55C - 1min
- 72C - 2min
I decreased the annealing phase from 60C to 55C from my standard program.
Results
<html><img src="http://dl.dropbox.com/u/8168852/IMG_20110106_101826.jpg" height="300px" width="400px"/></html>
Via Dropbox and my Droid.
So as you can barely tell in the above image, the reaction was successful in all 5 reactions. After cleanup I labeled the tube 1.1kb stretch, 1/6/11, ~400nM. I also attached a QR code that links to this page to the tube. In order to use, remove code via tape, scan code, click link. Using the same tape reattach the qr code to the tube.
Final concentration after PCR cleanup and nanodrop: 281.4ng/ul --> 383nM
