Ellington A, Pollard J D
University of Texas, Austin, Texas, USA.
Curr Protoc Mol Biol. 2001 May;Chapter 2:Unit2.12. doi: 10.1002/0471142727.mb0212s42.
Cloning vectors derived from filamentous phage are extremely useful because they allow cloned DNA to be isolated as either single- or double-stranded DNA. This unit contains protocols for preparing both forms of DNA and for characterizing inserts in M13-derived vectors. A protocol is also presented for preparing single-stranded DNA from plasmids using superinfection with helper phage. This method is advantageous because it allows cloned DNA to be maintained in the form of a plasmid while permitting single-stranded DNA to be isolated for DNA sequencing.
源自丝状噬菌体的克隆载体极其有用，因为它们能使克隆的DNA以单链或双链DNA的形式分离出来。本单元包含制备这两种DNA形式以及鉴定M13衍生载体中插入片段的实验方案。还介绍了一种使用辅助噬菌体超感染从质粒制备单链DNA的实验方案。这种方法具有优势，因为它能使克隆的DNA以质粒形式保存，同时又能分离出单链DNA用于DNA测序。
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