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限制性选择克隆:一种用于筛选重组DNA的简单通用方法。

Restriction selection cloning: a simple general method for the selection of recombinant DNA.

作者信息

Russek S J, Quirk J C, Farb D H

机构信息

Department of Pharmacology and Experimental Therapeutics, Boston University School of Medicine, MA 02118.

出版信息

Cell Mol Biol Res. 1993;39(2):177-82.

PMID:8220586
Abstract

It is well known that the ligation of two DNA fragments which are the product of digestion from different restriction enzymes will not lead to the regeneration of either of the original blunt end or cohesive end restriction sites. This property of sequence incompatibility for the original restriction enzyme can be exploited in a general cloning procedure for both PCR products and restricted DNAs. Restriction selection is particularly useful when cloning low abundance polymerase chain reaction (PCR) products and when cloning blunt ended DNA into reporter vectors that lack a method for the selection of recombinants.

摘要

众所周知,连接两个由不同限制酶消化产生的DNA片段,不会使原来的平端或粘性末端限制酶切位点再生。原始限制酶的这种序列不兼容性特性可用于PCR产物和经酶切的DNA的一般克隆程序。当克隆低丰度聚合酶链反应(PCR)产物,以及将平端DNA克隆到缺乏重组体选择方法的报告载体中时,限制酶选择特别有用。

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