PCR and Primer Dimer Question
Rod-Tweten at uokhsc.edu
Mon Oct 4 18:05:09 EST 1993
I was wondering if someone could tell me how much of a problem primer
dimers are in PCR. I am performing PCR mutagenesis on a gene that is AT
rich (about 70%) and it is almost impossible to get away from primer
dimers. For instance, the worst case is one where I have 8 bases out of
11 showing hybridization in a 22-mer. The bases are all A-T pairs.
Other examples would be 6 out of 7, again all A-T pairs. I realize that
primer dimers can kill a PCR reaction but can a hot start eliminate this
problem? Any help appreciated.
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