Drutsa V L, Kaberdin V R, Koroleva O N, Shilov I A
Bioorg Khim. 1991 Nov;17(11):1487-93.
A three primer variant of the earlier devised oligonucleotide-directed mutagenesis in plasmids is described, useful also for the fast cloning of single-stranded DNA products of the asymmetric polymerase chain reaction (PCR). Using this method for plasmid pHD-001-14-11, a 59 b. p. deletion and a 7 b. p. insertion were simultaneously introduced at 81% frequency, and the PCR-copied phage fd transcription terminator (26 b. p.) was inserted with the yield of 67%.
本文描述了一种改进的用于质粒的寡核苷酸定向诱变的三引物变体方法,该方法也可用于不对称聚合酶链反应(PCR)单链DNA产物的快速克隆。使用此方法对质粒pHD - 001 - 14 - 11进行操作,可同时以81%的频率引入59个碱基对的缺失和7个碱基对的插入,并且以67%的产率插入PCR复制的噬菌体fd转录终止子(26个碱基对)。