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Home Q & A Board Biology-Related Homework Help Cell Biology
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allan2009 allan2009
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13 years ago
In most PCR protocols, the primer pair need only be around 20 nucleotides each in length and annealing temperature is usually 55C; but in site-directed mutagenesis, the primers are often much longer with lengths from 25 to 45 nucleotides and the annealing temperature is around 65C. Two reasons why the primers need to be longer for site-directed mutagenesis?
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Educator
13 years ago
I know that longer primers increases the likelihood of secondary structure formation and it affects the efficiency of the mutagenesis reaction, but I'm not sure why. Moreover, longer primers also enhances the probability of non-specific reactions.
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