Nisson P E, Watkins P C, Krizman D B
Life Technologies, Gaithersburg, Maryland, USA.
Curr Protoc Hum Genet. 2001 May;Chapter 6:Unit 6.1. doi: 10.1002/0471142905.hg0601s03.
Exon trapping is an RNA polymerase chain reaction (PCR) method to clone expressed sequences or exons directly from mammalian genomic DNA. The basic protocol in this unit describes the method for trapping internal exons from cosmid clones and the second basic protocol describes trapping of 3 terminal exons. An describes 3 terminal exon trapping, which avoids subcloning of target DNA by ligating it to the vector for direct transfection. A describes a rapid cloning procedure using uracil DNA glycosylase.
外显子捕获是一种从哺乳动物基因组DNA中直接克隆表达序列或外显子的聚合酶链反应（PCR）方法。本单元中的基本方案描述了从黏粒克隆中捕获内部外显子的方法，第二个基本方案描述了3'末端外显子的捕获。方案A描述了3'末端外显子捕获，它通过将靶DNA连接到载体上进行直接转染，从而避免了靶DNA的亚克隆。方案B描述了一种使用尿嘧啶DNA糖基化酶的快速克隆程序。
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