Digestion of DNA with restriction endonucleases.
Digestion of DNA with restriction endonucleases.
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DOI：
10.1002/0471142727.mb0301s31
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发表时间：
2001-05-01
影响因子：
--
通讯作者：
Grossmann, B
中科院分区：
文献类型：
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作者：
Bloch, K D;Grossmann, B
限制性内切核酸酶识别短DNA序列并在识别序列内或邻近识别序列的特定位点切割双链DNA。限制性内切酶将DNA切割成离散片段是分子生物学中最基本的程序之一。本单元介绍的第一种方法是用单一限制性内切酶切割单一DNA样品。还描述了这种技术的一些常见应用。这些包括用多于一种内切核酸酶消化给定的DNA样品，用相同的内切核酸酶消化多个DNA样品，以及部分消化DNA使得切割仅发生在限制性位点的子集处。甲基化特定的DNA序列，并保护他们从限制性内切酶切割的协议也提出。在本单元的最后给出了一系列描述限制性内切酶及其特性的表格（包括有关识别序列、产生的末端类型、缓冲液条件和热灭活条件的信息）。
Restriction endonucleases recognize short DNA sequences and cleave double-stranded DNA at specific sites within or adjacent to the recognition sequences. Restriction endonuclease cleavage of DNA into discrete fragments is one of the most basic procedures in molecular biology. The first method presented in this unit is the cleavage of a single DNA sample with a single restriction endonuclease. A number of common applications of this technique are also described. These include digesting a given DNA sample with more than one endonuclease, digesting multiple DNA samples with the same endonuclease, and partially digesting DNA such that cleavage only occurs at a subset of the restriction sites. A protocol for methylating specific DNA sequences and protecting them from restriction endonuclease cleavage is also presented. A collection of tables describing restriction endonucleases and their properties (including information about recognition sequences, types of termini produced, buffer conditions, and conditions for thermal inactivation) is given at the end of the unit.