Atreya C D, Atreya P L, Pirone T P
Department of Plant Pathology, University of Kentucky, Lexington 40546.
Biochem Biophys Res Commun. 1990 Dec 31;173(3):1344-6. doi: 10.1016/s0006-291x(05)80935-0.
Polymerase chain reaction (PCR) based DNA amplification has replaced many time-consuming protocols in molecular biology. Here we describe a simple strategy to quickly select deletion mutants based on PCR methodology which then can be confirmed by nucleotide sequencing. A forward PCR primer is designed in such a way to recognize only the wild type sequences in the amplification reaction and thus a negative selection identifies the deletion in the samples.
基于聚合酶链反应(PCR)的DNA扩增已取代了分子生物学中许多耗时的实验方案。在此,我们描述了一种基于PCR方法快速筛选缺失突变体的简单策略,随后可通过核苷酸测序进行确认。正向PCR引物的设计方式使得其在扩增反应中仅识别野生型序列,因此通过负向筛选可鉴定样品中的缺失情况。