Site-specific mutagenesis in cells with normal DNA repair systems: transitions produced from DNA carrying a single O(6)-alkylguanine

在具有正常DNA修复系统的细胞中进行位点特异性诱变:由携带单个O(6)-烷基鸟嘌呤的DNA产生的转换

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Abstract

A simple and efficient method utilizing in vivo recombination to create recombinant libraries incorporating the products of PCR amplification is described. This will be especially useful for generating large pools of randomly mutagenized clones after error-prone PCR mutagenesis. Here we investigate various parameters to optimize this approach and we demonstrate that as little as 1 pmole of PCR fragment can generate a library with greater than 104 clones in a single transformation without ligation.

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