SSR标记需要特异引物进行扩增，因此SSR引物开发是一个关键问题。本文利用磁珠富集文库法来开发北柴胡SSR引物。以百花山柴胡（Bupleurum chinense DC. f. octoradiatum）为试材，首先将基因组DNA酶切后进行选择性扩增，然后用生物素标记的(AG)15、(AC)15和(MAB)12混合探针与其杂交，杂交片段与包被在磁珠表面的链亲和素（streptavidin）结合，经过一系列洗涤后，含有SSR的杂交片段通过PCR扩增得到双链目的片段，将这些片段连接到T载体上，转化大肠杆菌TOP10感受态细胞。经PCR法检测，最后对100个克隆进行测序,利用软件SSRHunter对序列进行微卫星统计，发现含有微卫星序列80个，大部分为（AC/TG）n、（TC/AG）n类型；其中有27个适合于设计引物。应用Primer Premier 5.0和在线设计软件Primer 3.0进行引物设计，得到27对引物，合成其中的8对，3对扩增出了预期长度的片段。本实验经关键步骤修正，建立了适合于开发柴胡SSR引物的FIASCO方法，为进一步分离大量北柴胡多态性卫星标记奠定了技术基础。

SSR marker needs specific primers for amplification, so the SSR primer development is an important problem.The objective of this work was to develop new SSR primers for Bupleurum Chinnese DC.. DNA of Bupleurum chinense DC. f. octoradiatum was used as materials. After enzmyed and selective amplification, the SSR were enriched by hybridizing (AG)15, (AC)15 and (MAB)12 probes with AFLP fragments and captured by magnetic beads coated with streptavidin. After washing to remove the non-hybridized fragments, the eluted single-stranded DNAs were enriched from preamplified AFLP fragments. Those became double-stranded by PCR amplification, were ligated into plasmid T vector, and then transformed into E. coli Top10 cells to produce a SSR-enriched library. After PCR screening, one hundred positive clones were sequenced. Through SSRHunter analysis, eighty sequences containing SSR were obtained. Most of them are abundant in（AC/TG）n、（TC/AG）n, and twenty seven are fit for primer designing. Twenty seven primers were designed using Primer Premier 5.0 or online Primer 3.0, and eight of them were synthesized. After PCR, three of the eight primers specifically produced fragments with the expected sizes. Therefore, the detailed protocols for the development of Bupleurum SSR primers by FIASCO method with modification has been established. It was proved to be a reliable method for isolating abundant SSRs for Bupleurum chinense DC.