ISSR-PCR和链亲和素磁珠吸附法开发白菜SSR引物  

SSR Primers Development in Non-heading Chinese Cabbage by ISSR-PCR and Streptavidin-coated Magnetic Beads Adsorption

作　　者：崔秀敏[1] 侯喜林[1] 董玉秀[2] 

机构地区：[1]南京农业大学作物遗传与种质创新国家重点实验室,南京210095 [2]山东农业大学生命科学院,泰安271018

出　　处：《园艺学报》2006年第1期155-157,共3页Acta Horticulturae Sinica

基　　金：国家‘863’计划项目(2003AA207120)

摘　　要：分别利用ISSR-PCR和链亲和素磁珠吸附法分离白菜基因组微卫星,共得到41对SSR引物。ISSR-PCR法运用巢式PCR,分别设计微卫星两侧引物IP2和IP3,SSR引物得率为12%。磁珠吸附法首先利用生物素标记的SSR探针与文库杂交,链亲和素磁珠吸附富集含SSR的片段。然后用接头引物扩增,克隆。根据测序结果,设计SSR两侧引物PL和PR,引物得率为9.6%。Two methods of ISSR-PCR and streptavidin-coated magnetic beads adsorption were used to isolate microsatellites from genomic DNA of non-heading Chinese cabbage （Brassica campestris ssp. chinensis） and a total of 41 SSR primers were successfully developed in this study. SSR primers （IP2 and IP3 ） were designed on the base of the determined terminal sequence of nested PCR products in the ISSR-PCR method with the efficiency of 12% for SSR primer development. As to the method of streptavidin-coated magnetic beads adsorption, firstly the biotin-labeled SSR probe was hybridized with digested genomic DNA and SSR fragments were enriched by absorption with strepavidin-coated magnetic beads and then they were amplified, cloned and sequenced. On the base of the sequence flanking microsatellites SSR primers （ PL and PR） were designed with an efficiency of 9.6%.

关 键 词：白菜 微卫星 ISSR-PCR 链亲和素磁珠吸附 SSR引物 

分 类 号：S634.3[农业科学—蔬菜学]