磁珠富集法制备莲藕基因组的微卫星分子标记  

Microsatellite Molecular Markers Enrichment and Isolation by Magnetic Beads （ FIASCO protocol） in Genome of Lotus（ Nelumbo nucifera Gaertn. ）

作　　者：潘磊[1] 郑鹏[1] 徐杰[1] 全志武[1] 李双梅 刘宏高[1] 柯卫东 丁毅[1] 

机构地区：[1]武汉大学生命科学学院,武汉大学植物发育生物学教育部重点实验室,430072 [2]国家种质武汉水生蔬菜资源圃,武汉430065

出　　处：《中国蔬菜》2007年第B08期7-13,共7页China Vegetables

基　　金：科技攻关计划项目（20062002043） 致谢：中国科学院水生生物研究所童金苟研究员给予技术方法上的帮助,在此感谢.

摘　　要：采用磁珠富集法FIASCO，设计生物素标记的寡核苷酸Biotin-（AG）13作探针，分离莲藕（Nelumbo nucifera Gaertn．）的微卫星分子标记，从所获得的阳性克隆中选取42个测序，结果36个克隆（85．0％）含有微卫星序列，其中23个（63．9％）重复数在10以上，19个（52．8％）为完美型。设计并且合成24对微卫星引物，经筛选其中16对为有效引物（66．7％），能扩增出多态性条带。Microsatellite enrichment by magnetic beads, fast isolation by the AFLP of sequences containing repeats （FIASCO protocol）, was used to isolate and characterize microsatellite molecular markers of Nelumbo nucifera Gaertn. by Biotin - （AG） 13 as probe. 42 positive clones were sequenced. 36 of the sequenced clones（ about 85 % ） contained SSR locus, among which 63.9 % （23 clones） microsatellites repeated more than 10 times and 52.8 % （ 19 clones） microsatellites were perfect repeats. Then,24 pairs of primers were designed,and 16 of them（66.7 % ）were effective primer pairs,by which the polymorphism bands could be amplified. These showed microsatellite enrichment by magnetic beads is an effective method to develop microsatellites. At the same time ,the microsatellites obtained can offer useful genetic markers for studying Nelumbo nucifera in the future.

关 键 词：莲藕 磁珠富集法 微卫星 基因组 

分 类 号：S512.1[农业科学—作物学]