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From 2013.igem.org

June 24th 2013

Patrick - Set up a 25 uL Q5 master mix PCR reaction to extract RcoM from the Azo gDNA. Used RcoM_F and RcoM_R with a Tm of 52 and 56, an elongation time of 1:15 minutes and 32 cycles. This is the Tm established by our analysis with APE when creating the primers. The Tm from the sides of the primer tubes would suggest a Tm of 60 C but this is taking into account the full length of the primer (including the overhang). RcoM is 495 bp and the master mix has an extension of 30 seconds / kb. 30-35 cycles is recommended for gDNA amplification. I used 2 uL of template DNA (~50 ng). Tomorrow Eric will run a gel and isolate HemAT.

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