荧光标记STR复合扩增检验系统的研究

本研究以化学合成的荧光标记引物建立了STR复合扩增体系,体系I包括4个STR基因座和牙釉蛋白基因,体系Ⅱ包括4个STR基因座,体系Ⅲ包括3个STR基因座.制备了体系I各基因座的等位基因标准对照,测定了各等位基因长度.对代表性等位基因进行了序列测定,确定了等位基因重复单位的重复数目及测量长度与等位基因命名的对应关系,对各基因座等位基因进行了标准命名.建立了复合扩增检验系统等位基因自动分析命名的模板.三个复合扩增检验体系的累积随机匹配概率为8.3×10-3,为遗传学分析建立了实用的检验系统.

MULTIPLEX SETS FOR THE AMPLIFICATION OF FLUORESCENTLY TAGGED SHORT TANDEM REPEAT LOCI

Multiplex PCR amplification system s were developed using synthesized f luorescently tagged primers of11well -characterized short tandem repeat loci and the X -Y homologous am elogenin gene.Multiplex configura tions contained a mixture of either four or three separate primer pairs for simu ltaneous amplification of multiple loci in a single reaction tube.A mixture of allelic l adders consisting of all the common a lleles for the respective loci in mul tiplex I was prepared,and the window for each all ele was set based on the range of autom atic size calling against the GS500i nternal size markers.Reprensentative alle le of each STR locus has been sequenced in order to establish the repeat uni t sequence and assign allelic designation acco rding to the basic recommendations o f the DNA commission of the Internati onal So-ciety of Forensic Haemogenetics.Ra nges and nomenclature were programmed into the ABD GENOTYPER TM software,enabling automatic allele designation to be made.