Ledwith B J, Manam S, Nichols W W, Bradley M O
Merck Sharp and Dohme Research Laboratories, West Point, PA 19486.
Biotechniques. 1990 Aug;9(2):149-52.
DNA fingerprints are generated using probes that hybridize to hypervariable minisatellites, also known as variable number tandem repeat loci. Cloned minisatellites have served as the predominant source of DNA fingerprinting probes. A short segment within the repeat units of minisatellites, called the "core" sequence, is highly conserved within a family of related minisatellites, thereby allowing a single-cloned minisatellite to cross-hybridize to 20 to 40 other minisatellites. In this article, we describe a method for the synthetic preparation of polymeric core sequence probes for DNA fingerprinting. Unlike "monomeric" oligonucleotide probes, the polymeric probes mimic the tandem-repetitive structure of minisatellites, and thus each probe molecule can potentially form many sites of hybridization with a target minisatellite. The synthetic probes are cloned into plasmid DNA to provide a perpetual source of probe material.
DNA指纹图谱是使用与高变微卫星(也称为可变数目串联重复序列位点)杂交的探针生成的。克隆的微卫星一直是DNA指纹图谱探针的主要来源。微卫星重复单元内的一小段序列,称为“核心”序列,在相关微卫星家族中高度保守,因此单个克隆的微卫星可以与另外20至40个微卫星交叉杂交。在本文中,我们描述了一种用于DNA指纹图谱的聚合核心序列探针的合成制备方法。与“单体”寡核苷酸探针不同,聚合探针模仿微卫星的串联重复结构,因此每个探针分子可能与目标微卫星形成多个杂交位点。合成探针被克隆到质粒DNA中,以提供探针材料的永久来源。