Insertion and Deletion Mutagenesis by Overlap Extension PCR
Insertion and Deletion Mutagenesis by Overlap Extension PCR
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DOI：
10.1007/978-1-60761-652-8_10
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发表时间：
2010-01-01
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影响因子：
--
通讯作者：
Ryu, Wang-Shick
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文献类型：
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作者：
Lee, Jehan;Shin, Myeong-Kyun;Ryu, Wang-Shick
通过重叠延伸PCR的诱变已经成为产生突变（包括取代、插入和缺失）的标准方法。然而，建立的重叠PCR诱变在许多方面是有限的。特别是，很难使插入大于30 μ m，因为所有的序列改变必须嵌入引物内。在这里，我们描述了一种快速有效的方法，用于在DNA分子中的任何位置创建任何长度的插入或缺失。该方法是普遍适用的，因此代表了对现在广泛使用的重叠延伸PCR方法的显著改进。
Mutagenesis by the overlap extension PCR has become a standard method of creating mutations including substitutions, insertions, and deletions. Nonetheless, the established overlap PCR mutagenesis is limited in many respects. In particular, it has been difficult to make an insertion larger than 30 it, since all sequence alterations must be embedded within the primer. Here, we describe a rapid and efficient method for creating insertions or deletions of any length at any position in a DNA molecule. This method is generally applicable, and therefore represents a significant improvement to the now widely used overlap extension PCR method.