An efficient PCR mutagenesis strategy without gel purification step that is amenable to automation
An efficient PCR mutagenesis strategy without gel purification step that is amenable to automation
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DOI：
10.1093/nar/24.16.3276
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发表时间：
1996-08-15
影响因子：
14.9
通讯作者：
KandelsLewis, S
中科院分区：
文献类型：
--
作者：
Seraphin, B;KandelsLewis, S
我们在这里描述了一种改进的巨引物PCR诱变策略。通常使用的繁琐的凝胶纯化步骤可以通过用限制性内切酶适当地切割第一和第二DNA模板并从第一次PCR反应中酶切去除剩余的引物来省略。我们证明了这种改进的方法是可重复的和高效的。此外，该方法适用于自动化，因为所有步骤现在都在反应管中进行。
We describe here an improved megaprimer PCR mutagenesis strategy. The cumbersome gel purification step that is usually used can be omitted by appropriately cleaving the first and second DNA templates with restriction enzymes and enzymatically removing remaining primers from the first PCR reaction. We show that this improved procedure is reproducible and highly efficient. Furthermore this method is suitable for automation because all the steps are now carried out in reaction tubes.