Efficiency of the Polymerase Chain Reaction.

Efficiency of the Polymerase Chain Reaction.
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DOI:
10.1016/j.ces.2010.05.046
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发表时间:
2010-09-01
影响因子:
4.7
通讯作者:
Viljoen HJ
Viljoen HJ
中科院分区:
工程技术2区
文献类型:
--
作者:
Booth CS;Pienaar E;Termaat JR;Whitney SE;Louw TM;Viljoen HJ

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聚合酶链反应(PCR)在生物化学和分子生物学中有着广泛的应用,如基因表达研究、突变检测、法医学分析和病原体检测等。越来越多的定量真实的时间PCR用于评估来自总产量的拷贝数。在这项研究中,产量作为几个过程的函数进行分析:(1)在变性步骤中发生模板和聚合酶的热损伤,(2)引物和模板之间存在竞争,以退火或形成dsDNA,(3)聚合酶结合退火产物(引物/ssDNA),以形成三元复合物和(4)三元复合物的延伸。为每个过程的效率提供了明确的表达式,因此反应条件可以直接与总产率相关联。提供的例子中,不同的过程发挥产量限制的作用。该分析将使研究人员对控制反应的因素有独特的理解,并有助于解释实验结果。
The polymerase chain reaction (PCR) has found wide application in biochemistry and molecular biology such as gene expression studies, mutation detection, forensic analysis and pathogen detection. Increasingly quantitative real time PCR is used to assess copy numbers from overall yield. In this study the yield is analyzed as a function of several processes: (1) thermal damage of the template and polymerase occurs during the denaturing step, (2) competition exists between primers and templates to either anneal or form dsDNA, (3) polymerase binding to annealed products (primer/ssDNA) to form ternary complexes and (4) extension of ternary complexes. Explicit expressions are provided for the efficiency of each process, therefore reaction conditions can be directly linked to the overall yield. Examples are provided where different processes play the yield-limiting role. The analysis will give researchers a unique understanding of the factors that control the reaction and will aid in the interpretation of experimental results.
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发表时间: 1988-09-29
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