Quantitative PCR

Rajesh R. Naik rn2c+ at andrew.cmu.edu
Fri Mar 10 12:01:13 EST 1995


Hi,

I am attempting to develop competitive I have cloned the internal
standard DNA into a pGEM vector in order to facilitate in-vitro
transcription. Internal control DNA differs from the wild type target
DNA by 350bp. I would like to do quantitative PCR so that the
competition between WT and the internal standard start at the reverse
transcription step.

My question is, how does one calculate copy numbers/cell( of the gene of
interest) and be able to plot the standard against the copy number, to
eventually estimate the copy number of the target DNA 



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