Chong S, Garcia G A
University of Michigan, Ann Arbor 48109-1065.
Biotechniques. 1994 Oct;17(4):719-20, 722, 724-5.
The sequential addition of primers in a PCR enables one to preferentially amplify one of the two strands of a heteroduplex DNA template. This serves as the basis for a novel site-directed mutagenesis technique involving a heteroduplex DNA template that has been generated from a single-stranded, wild-type template and one or more mutagenic oligonucleotides. This preferential PCR method yields a mutation efficiency greater than 90% (consistent with the theoretical estimate for the method). The ability to generate multiple mutants also enables the screening of potential mutants by restriction endonuclease digestion.
在聚合酶链式反应（PCR）中依次添加引物，能够使人们优先扩增异源双链DNA模板两条链中的一条。这为一种新型的定点诱变技术奠定了基础，该技术涉及由单链野生型模板与一种或多种诱变寡核苷酸生成的异源双链DNA模板。这种优先PCR方法产生的突变效率大于90%（与该方法的理论估计一致）。生成多个突变体的能力还使得通过限制性内切酶消化来筛选潜在突变体成为可能。
