Gibbs R A, Nguyen P N, Caskey C T
Institute for Molecular Genetics, Baylor College of Medicine, Houston, TX 77030.
Nucleic Acids Res. 1989 Apr 11;17(7):2437-48. doi: 10.1093/nar/17.7.2437.
Synthetic DNA oligonucleotides can serve as efficient primers for DNA synthesis even when there is a single base mismatch between the primers and the corresponding DNA template. However, when the primer-template annealing is carried out with a mixture of primers and at low stringency the binding of a perfectly matched primer is strongly favored relative to a primer differing by a single base. This primer competition is observed over a range of oligonucleotide sizes from twelve to sixteen bases and with a variety of base mismatches. When coupled with the polymerase chain reaction, for the amplification of specific DNA sequences, competitive oligonucleotide priming provides a simple general strategy for the detection of single DNA base differences.
合成DNA寡核苷酸即使在引物与相应DNA模板之间存在单个碱基错配时，也可作为DNA合成的有效引物。然而，当用引物混合物在低严谨性条件下进行引物-模板退火时，与相差单个碱基的引物相比，完全匹配的引物的结合受到强烈偏好。在12至16个碱基的一系列寡核苷酸大小范围内以及各种碱基错配情况下，均观察到这种引物竞争现象。当与聚合酶链反应相结合用于特定DNA序列的扩增时，竞争性寡核苷酸引物引发为检测单个DNA碱基差异提供了一种简单通用的策略。
