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Mitch2481 Mitch2481
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14 years ago
In most PCR amplification reactions, the primer pair is "bookended" at either end of the template antiparallel strands that needto be amplified. However, in site-directed mutagenesis, the primers are actually "back-to-back". Why is this different arrangment of the primers necessary?
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wrote...
14 years ago
In PCR, primer pairs are bookended so that if the gene is present, these primers will bind to the corresponding target sequences, thereby bookending the gene of interest. In site-directed mutagenesis, the basic procedure requires the synthesis of a short DNA primer containing the desired base change. Its primer is back-to-back because it is the only way to hybridize with a broken (mutated), single-stranded DNA containing the gene of interest.

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14 years ago
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