Fisher P J, Gardner R C, Richardson T E
New Zealand Forest Research Institute Ltd, Rotorua.
Nucleic Acids Res. 1996 Nov 1;24(21):4369-71. doi: 10.1093/nar/24.21.4369.
Microsatellites are widely used as genetic markers because they are co-dominant, multiallelic, easily scored and highly polymorphic. A major drawback of microsatellite markers is the time and cost required to characterise them. We have developed a novel technique to reduce this cost by producing a microsatellite-rich PCR profile from genomic DNA which was cloned to yield a genomic library enriched for microsatellites. Sequence data and subsequent allele scoring within pedigrees revealed that these microsatellites retained their original repeat length and segregated normally. This technique permits genomic amplification with only one specific primer. Together with enrichment, the savings in primer costs reduces the cost of microsatellite characterisation considerably.
微卫星广泛用作遗传标记,因为它们是共显性的、多等位基因的、易于计分且高度多态的。微卫星标记的一个主要缺点是表征它们所需的时间和成本。我们开发了一种新技术,通过从基因组DNA产生富含微卫星的PCR图谱来降低成本,该基因组DNA被克隆以产生富含微卫星的基因组文库。系谱内的序列数据和随后的等位基因计分表明,这些微卫星保留了其原始重复长度并正常分离。该技术仅用一种特异性引物即可进行基因组扩增。连同富集一起,引物成本的节省大大降低了微卫星表征的成本。