Molecular Cloning Fourth Edition, A Laboratory Manual, by Michael R. Green and Joseph Sambrook

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Hot Start PCR

(Protocol summary only for purposes of this preview site)

Mispairing of primers, which occurs at suboptimal annealing temperatures, leads to the synthesis of nonspecific PCR products. Off-target amplification can become a serious problem when PCRs are performed with low concentrations of a complex template, such as mammalian genomic DNA template (Chou et al. 1992). In reactions of this type, the possibility for mispriming is increased by the elevated ratio of primer:template DNA.


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(Limited time special offer.) Molecular Cloning Fourth Edition, A Laboratory Manual, by Michael R. Green and Joseph Sambrook
Molecular Cloning Fourth Edition, A Laboratory Manual, by Michael R. Green and Joseph Sambrook

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