中文摘要：

报道了一种简便高效克隆微卫星序列的方法。这一方法的实施步骤包括小片段基因组文库构建、三螺旋捕获、磁珠分离和PCR扫描。草鱼AG重复文库的冗余率为7.2%,富集效率为60.25倍,PCR扫描的准确率达100%,采用这一方法克隆到的AG重复序列中完美型、非完美型和混合型的比例分别为66.87%、17.17%和15.96%,不间断重复序列长度大于30 bp的微卫星座位占51%。34个微卫星座位中有25个表现出多态性,PIC值0.50—0.91,进一步分析表明AG重复序列的多态信息含量（PIC）与不间断重复序列长度相关,不间断重复序列长度大于30 bp的座位表现出丰富的多态性。较高的富集率和有效引物获得率证明此方法在分离草鱼微卫星中的成功应用,并将为草鱼养殖品系的优化、遗传多样性的检测及遗传图谱的构建等打下基础。

英文摘要：

A simple and efficient method for isolating microsatellites was described.The approach involves small-insert genomic library construction,triplex affinity capture,magnetic separation,and PCR screen.The redundancy of grass carp（AG）-enriched library was 7.2%,the enrichment rate was 68.8-fold,and the accuracy rate of PCR screening was 100%.In（AG） repeats we obtained,perfect,imperfect and compound type occupied 66.87%,17.17% and 15.96% respectively,and uninterrupted repeat sequences longer than 30 bp accounted for 51%.Polymorphism analysis of 34（AG） repeat sequence loci indicated that 25 repeats of them were polymorphic with 0.50—0.91 PIC.Further analysis revealed that the informativeness of the（AG） repeat sequence loci increased with an increasing number of repeats and uninterrupted repeats longer than 30 bp exhibited abundant polymorphisms.High enrichment rate and informative microsatellites acquisition rate demonstrated the successful application of Triple-Helix-Mediated affinity capture in the isolation of microsatellites in grass carp,and this will build a good foundation for the improvement of breeding lines,genetic diversity detection and genetic map construction in grass carp.