Department of Molecular Biology & Genetics, Aarhus University, C.F. Møllers Allé 3, 8000 Aarhus C, Denmark.
Biotechniques. 2020 Jun;68(6):345-348. doi: 10.2144/btn-2019-0104. Epub 2020 May 6.
We describe the application of simple cloning by prolonged overlap extension for multiple site-directed mutagenesis in the same plasmid. We show that it is possible to use this technique with very short PCR templates. The technique is ideally suited for the generation of longer donor DNA sequences for CRISPR/Cas9-mediated homologous repair.
我们描述了一种通过延长重叠延伸进行多位点定点突变的简单克隆方法在同一质粒中的应用。我们表明，该技术可以使用非常短的 PCR 模板。该技术非常适合用于生成更长的供体 DNA 序列，用于 CRISPR/Cas9 介导的同源修复。
