Rashtchian A
Life Technologies Inc, Gaithersburg.
Curr Opin Biotechnol. 1995 Feb;6(1):30-6. doi: 10.1016/0958-1669(95)80006-9.
Use of the polymerase chain reaction (PCR) has become increasingly widespread in virtually all aspects of molecular biology. Recently, novel ligation-independent methods have been developed for the cloning of DNA fragments amplified using PCR. Ligation-independent cloning utilizing the enzyme uracil DNA glycosylase (termed UDG cloning) provides an efficient method for gene cloning and recombinant PCR. This technology is now being applied to site-directed mutagenesis, the generation of nested deletions, and the engineering of novel gene constructs. The ease and flexibility of this methodology, combined with PCR amplification, simplify gene cloning and engineering techniques.
聚合酶链反应（PCR）在分子生物学的几乎所有领域中的应用都越来越广泛。最近，已经开发出了用于克隆通过PCR扩增的DNA片段的新型非连接方法。利用尿嘧啶DNA糖基化酶的非连接克隆（称为UDG克隆）为基因克隆和重组PCR提供了一种有效的方法。该技术目前正应用于定点诱变、嵌套缺失的产生以及新型基因构建体的工程改造。这种方法的简便性和灵活性，与PCR扩增相结合，简化了基因克隆和工程技术。
