Amplification of closed circular DNA in vitro
Amplification of closed circular DNA in vitro
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DOI：
10.1093/nar/26.4.1126
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发表时间：
1998-02-15
影响因子：
14.9
通讯作者：
Ruffner, DE
中科院分区：
文献类型：
--
作者：
Chen, ZD;Ruffner, DE
聚合酶链反应是一种用于体外扩增核酸的强大技术，该反应产生线性产物，到目前为止，封闭的环状产物还不可能，因为许多DNA分子的复制能力形式是封闭的环状形式，将封闭的环状DNA扩增为封闭的环状分子是不可能的。到目前为止，这些分子只能在适当的宿主细胞中在体内扩增，在这里，我们描述了一种体外程序，连接期间扩增（LDA），用于使用序列特异性引物选择性扩增封闭环状DNA。LDA可用于定点诱变、突变检测、DNA修饰、DNA文库筛选和环状DNA生产。
The polymerase chain reaction is a powerful technique used to amplify nucleic acids in vitro, The reaction produces linear products, and as of yet, closed circular products have not been possible, Since the replicatively competent form of many DNA molecules is the closed circular form, it would be adventitious to amplify closed circular DNA as closed circular molecules. Until now, these molecules could only be amplified in vivo in appropriate host cells, Here, we describe an in vitro procedure, ligation-during-amplification (LDA), for selective amplification of closed circular DNA using sequence-specific primers. LDA is useful for site-directed mutagenesis, mutation detection, DNA modification, DNA library screening and circular DNA production.