Mitsuhashi M
Hitachi Chemical Research Center, Irvine, CA 92715, USA.
J Clin Lab Anal. 1996;10(5):285-93. doi: 10.1002/(SICI)1098-2825(1996)10:5<285::AID-JCLA9>3.0.CO;2-7.
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.
设计最佳聚合酶链反应（PCR）引物序列是实现具有灵敏、特异且各检测间可重复结果的成功PCR的关键因素之一。在本综述中，总结了PCR引物序列的所有要求，如位置、扩增子大小、引物长度、核苷酸组成、解链温度（Tm）、3'端杂交强度和频率、发夹结构形成能、引物间相互作用、特异性以及与交叉杂交序列错配的位置。本报告还讨论了如何探索这些各类信息以用于更先进的PCR应用，包括巢式PCR、多重PCR、竞争性PCR、长片段PCR、点突变检测、简并引物以及PCR克隆。
