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

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Touchdown PCR

(Protocol summary only for purposes of this preview site)

The specificity of conventional PCRs decreases as the sequence complexity of the template DNA increases. The greater the complexity, the greater is the chance that the primers will bind promiscuously to sequences other than the intended target. The number of mispriming events can be reduced by optimizing the concentrations of the components of the PCR, in particular the concentrations of Mg2, primers, dNTPs, and template. Further minimization of off-target amplification can be obtained by using the most stringent annealing conditions that permit efficient, specific annealing of the oligonucleotide primers to their targets.


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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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