Heath D D, Iwama G K, Devlin R H
Fisheries and Oceans Canada, West Vancouver Laboratory, BC.
Nucleic Acids Res. 1993 Dec 11;21(24):5782-5. doi: 10.1093/nar/21.24.5782.
The use of genomic DNA-based techniques in ecological and evolutionary studies has been limited by the availability of suitable probes for species of interest due to the technical difficulty of isolating and applying such probes. We have developed a simple technique that directs polymerase chain reaction (PCR) amplification to regions rich in variable number of tandem repeats (VNTRs). By using published VNTR core sequences as primers in PCRs, fragments were amplified that showed little variation within a species, but did show differences between species. When the amplified fragments were used as probes with genomic DNA Southern blots they produced hypervariable single-locus or few-locus patterns in fish, birds, and humans. We have named this procedure as Directed Amplification of Minisatellite-region DNA (DAMD).
基于基因组DNA的技术在生态和进化研究中的应用一直受到限制，原因是由于分离和应用此类探针存在技术困难，难以获得针对感兴趣物种的合适探针。我们开发了一种简单的技术，可将聚合酶链反应（PCR）扩增导向富含可变串联重复序列（VNTR）的区域。通过使用已发表的VNTR核心序列作为PCR引物，扩增出的片段在物种内显示出很少的变异，但在物种间确实存在差异。当将扩增片段用作基因组DNA Southern杂交的探针时，它们在鱼类、鸟类和人类中产生了高变单基因座或少数基因座模式。我们将此程序命名为小卫星区域DNA的定向扩增（DAMD）。
