当前位置: 首页 > 文章 > 利用磁珠富集法开发花榈木微卫星引物 华中农业大学学报 2010,29 (5) 99-103
Position: Home > Articles > Development of Microsatellite Primers in Ormosia henryi Prain by Magnetic Beads Enrichment Journal of Huazhong Agricultural University 2010,29 (5) 99-103

利用磁珠富集法开发花榈木微卫星引物

作  者:
胡磊;高丽;杨波
单  位:
中国科学院武汉植物园
关键词:
花榈木;微卫星标记;磁珠富集;多态性
摘  要:
将花榈木(Ormosia henryiPrain)基因组DNA用MseⅠ酶切后,连接相应的接头,PCR扩增后回收500~1000bp片段,与用生物素标记的微卫星探针(GA)12,(AAG)8杂交后,运用磁珠富集含有微卫星序列的DNA片段。获得的序列经PCR扩增,连接载体pMD19-T,构建富含微卫星序列的小片段插入文库;用PCR法筛选文库,获得78个阳性克隆,经测序分析得到24个微卫星序列,成功设计出9对引物。
译  名:
Development of Microsatellite Primers in Ormosia henryi Prain by Magnetic Beads Enrichment
作  者:
HU Lei 1,2 GAO Li1 YANG Bo11.Wuhan Botanical Garden,Chinese Academy of Sciences,Wuhan 430074,China;2.Graduate University of Chinese Academy of Sciences,Beijing 100049,China
关键词:
Ormosia henryi Prain;microsatellite markers;magnetic beads enrichment;polymorphic
摘  要:
The genomic DNA of Ormosia henryi Prain was digested by MseⅠand ligated with adapters.After PCR amplification,the fragments ranging from 500 to 1 000 bp were isolated,purified and hybridized with biotin-labeled probes(GA)12 and(AAG)8.Fragments containing simple sequence repeats(SSR)were enriched using magnetic beads coated with streptavidin,then cloned into pMD19-T vectors and transformed into E.coli to construct SSR genomic DNA library.78 positive clones were screened with PCR method.After sequenced,24 microsatellite sequences were identified and 9 pairs of SSR primers of Ormosia henryi Prain were developed successfully.

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