聚合酶
寡核苷酸
热启动PCR
脱氧核糖核酸
聚合酶
聚合酶链反应
分子生物学
DNA聚合酶
多重位移放大
底漆(化妆品)
聚合酶链反应优化
DNA
化学
底漆二聚体
变性(裂变材料)
色谱法
生物化学
生物
水热
DNA提取
多重聚合酶链反应
基因
有机化学
核化学
作者
Martha F. Kramer,Donald M. Coen
标识
DOI:10.1002/0471142727.mb1501s56
摘要
This unit describes a method for amplifying DNA enzymatically by the polymerase chain reaction (PCR), including procedures to quickly determine conditions for successful amplification of the sequence and primer sets of interest, and to optimize for specificity, sensitivity, and yield. The first step of PCR simply entails mixing template DNA, two appropriate oligonucleotide primers, Taq or other thermostable DNA polymerases, deoxyribonucleoside triphosphates (dNTPs), and a buffer. Once assembled, the mixture is cycled many times (usually 30) through temperatures that permit denaturation, annealing, and synthesis to exponentially amplify a product of specific size and sequence. The PCR products are then displayed on an appropriate gel and examined for yield and specificity. Recommended optimization conditions are included.
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