Technical report .2. Basic requirements for designing optimal PCR primers
Technical report .2. Basic requirements for designing optimal PCR primers
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DOI：
10.1002/(sici)1098-2825(1996)10:5
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发表时间：
1996-01-01
影响因子：
2.7
通讯作者：
Mitsuhashi, M
中科院分区：
文献类型：
--
作者：
Mitsuhashi, M
设计最佳的聚合酶链式反应(PCR)引物序列是成功的聚合酶链式反应(PCR)获得灵敏、特异和重复性结果的关键因素之一。本文综述了扩增片段的位置、大小、长度、核苷酸组成、Tm、3‘端杂交强度和频率、发夹形成能、引物间相互作用、特异性以及与杂交序列错配的位置等对扩增产物序列的要求。该报告还讨论了如何利用这些不同类型的信息来进行更高级的聚合酶链式反应应用，包括套式聚合酶链式反应、多重聚合酶链式反应、竞争性聚合酶链式反应、长聚合酶链式反应、点突变检测、简并引物和聚合酶链式反应克隆。(C)1996年Wiley-Liss，Inc.
Designing optimal polymerase chain reaction (PCR) primer sequences is one of the critical factors for successful PCR with sensitive, specific, and assay-to-assay reproducible results. In this review, all the requirements of PCR primer sequences are summarized, such as location, size of amplicon, length of primers, nucleotide composition, Tm, 3' terminal hybridization strength and frequency, hairpin formation energy, primer-to-primer interaction, specificity, and location of mismatches to sequences of cross-hybridization. The report also discusses how to explore these various types of information for more advanced PCR applications, which include nested PCR, multiplex PCR, competitive PCR, long PCR, point mutation detection, degenerate primers, and PCR cloning. (C) 1996 Wiley-Liss, Inc.