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单词 Touchdown PCR
释义

Touchdown PCR

中文百科

递减聚合酶链式反应 Touchdown polymerase chain reaction

(重定向自Touchdown PCR)

递减PCR(touchdown PCR)是一种PCR(聚合酶链式反应)方法,用来避免非特异性串行的扩增。PCR中引物的黏合温度(annealing temperature)决定了黏合的特异性,温度越高特异性越强,但过高则不能实现引物和模板的结合,而过低会产生大量非特异性产物。因此进行PCR时需要寻找合适的黏合温度。

递减PCR开始先设置一个比较高的黏合温度,每个循环黏合温度下降1℃,到一个较低温度以后每个循环的黏合温度保持不变直到结束。在刚下降到特异性结合可以进行的最高温度时,可以达到最高的特异性。这样在起初的几个循环中,特异性扩增串行可以相对非特异性串行多扩增若干倍,从而减轻非特异性扩增对结果的干扰。这种方法可用于省去多次试验最佳黏合温度的工作。

英语百科

Touchdown polymerase chain reaction 递减聚合酶链式反应

(重定向自Touchdown PCR)

The touchdown polymerase chain reaction or touchdown style polymerase chain reaction is a method of polymerase chain reaction by which primers will avoid amplifying nonspecific sequences. The annealing temperature during a polymerase chain reaction determines the specificity of primer annealing. The melting point of the primer sets the upper limit on annealing temperature. At temperatures just below this point, only very specific base pairing between the primer and the template will occur. At lower temperatures, the primers bind less specifically. Nonspecific primer binding obscures polymerase chain reaction results, as the nonspecific sequences to which primers anneal in early steps of amplification will "swamp out" any specific sequences because of the exponential nature of polymerase amplification.

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更新时间:2025/6/20 16:55:26