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Method for polymerase chain reactions with use of a DNA polymerase with proofreading properties

机译:使用具有校对特性的DNA聚合酶进行聚合酶链反应的方法

摘要

The present invention concerns a method for a polymerase chain reaction, in which a template nucleic acid, at least one primer, deoxyribonucleoside triphosphates as well as a DNA polymerase with proofreading activity are used. In addition, according to this invention, at least one target substrate is added to the polymerase chain reaction, whereby the efficiency of the DNA polymerase with proofreading activity is significantly increased. Any molecule that reduces or, in the optimal case, blocks the 3′,5′-exonuclease activity of the DNA polymerase used is suitable as target substrate. Technical solutions for the added substrate (target substrate) are in particular single stranded, linear oligonucleotides, hairpin oligonucleotides and RNA and DNA molecules. Furthermore, a kit is disclosed which comprise the required reagents for the implementation of the method according to the invention.
机译:本发明涉及一种用于聚合酶链式反应的方法,其中使用模板核酸,至少一个引物,脱氧核糖核苷三磷酸以及具有校对活性的DNA聚合酶。另外,根据本发明,将至少一种靶底物加入到聚合酶链反应中,从而显着提高了具有校对活性的DNA聚合酶的效率。降低或在最佳情况下阻断所用DNA聚合酶的3',5'-核酸外切酶活性的任何分子均适合作为靶底物。用于添加的底物(靶底物)的技术解决方案特别是单链,线性寡核苷酸,发夹寡核苷酸以及RNA和DNA分子。此外,公开了一种试剂盒,其包含用于实施根据本发明的方法所需的试剂。

著录项

  • 公开/公告号US9309567B2

    专利类型

  • 公开/公告日2016-04-12

    原文格式PDF

  • 申请/专利权人 HOLGER ENGEL;RALF PEIST;

    申请/专利号US20060540492

  • 发明设计人 RALF PEIST;HOLGER ENGEL;

    申请日2006-09-28

  • 分类号C12Q1/68;

  • 国家 US

  • 入库时间 2022-08-21 14:33:26

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