ISSR-PCR和链亲和素磁珠吸附法开发白菜SSR引物

园艺学报 ›› 2006, Vol. 33 ›› Issue (1): 155-157.

ISSR-PCR和链亲和素磁珠吸附法开发白菜SSR引物

崔秀敏¹;侯喜林^1*;董玉秀²

(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)

收稿日期:2005-03-07 修回日期:2005-06-17 出版日期:2006-02-25 发布日期:2006-02-25

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

Cui Xiumin¹;Hou Xilin^1*;and Dong Yuxiu²

(¹National Key Laboratory of Crop Genetics and Germplasm Enhancement, Nanjing Agricultural University, Nanjing 210095,China; ²College of Life Science, Shandong Agricultural University, Tai'an 271018, China)Abstract:

Received:2005-03-07 Revised:2005-06-17 Online:2006-02-25 Published:2006-02-25

摘要/Abstract

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%。

关键词: 白菜, 微卫星, ISSR-PCR, 链亲和素磁珠吸附, SSR引物

Abstract: 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 p robe was hybridized with digested genomic DNA and SSR fragments were enriched by absorption with strepavidin-coated magnetic beads and then theywere amp lified, cloned and
sequenced. On the base of the sequence flanking microsatellites SSR primers(P_L and P_R ) were designed with an efficiency of 9.6%.

Key words: Non-heading Chinese cabbage, Microsatellite, ISSR-PCR, Streptavidin-coated magnetic beads adsorption, SSR primer

崔秀敏;侯喜林;董玉秀. ISSR-PCR和链亲和素磁珠吸附法开发白菜SSR引物[J]. 园艺学报, 2006, 33(1): 155-157.

Cui Xiumin;Hou Xilin;and Dong Yuxiu. SSR Primers Development in Non-head ing Chinese Cabbage by ISSR-PCR and Streptavidin-coated Magnetic Beads Adsorption[J]. ACTA HORTICULTURAE SINICA, 2006, 33(1): 155-157.

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