| ISSR-PCR和链亲和素磁珠吸附法开发白菜SSR引物 |
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| 引用本文： | 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 |
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| 作者姓名： | 崔秀敏 侯喜林 董玉秀 |
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| 作者单位： | ( 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; 山东农业大学生命科学院, 泰安271018) |
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| 基金项目： | 国家‘863’计划项目(2003AA207120) |
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| 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%。
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| 关 键 词： | 白菜 微卫星 ISSR-PCR 链亲和素磁珠吸附 SSR引物 |
| 文章编号： | 0513-353X（2006）01-0155-03 |
| 收稿时间： | 2005-03-07 |
| 修稿时间： | 2005-03-072005-06-17 |
SSR Primers Development in Non-heading Chinese Cabbage by ISSR-PCR and Streptavidin-coated Magnetic Beads Adsorption |
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| Cui Xiumin,Hou Xilin,Dong Yuxiu. SSR Primers Development in Non-heading Chinese Cabbage by ISSR-PCR and Streptavidin-coated Magnetic Beads Adsorption[J]. Acta Horticulturae Sinica, 2006, 33(1): 155-157 |
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| Authors: | Cui Xiumin Hou Xilin Dong Yuxiu |
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| Affiliation: | (National Key Laboratory of Crop Genetics and Germplasm Enhancement, Nanjing Agricultural University, Nanjing 210095,China; College of Life Science, Shandong Agricultural University, Tai'an 271018, China)Abstract: |
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| 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(IP_2 and IP_3) 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(P_L and P_R) were designed with an efficiency of 9.6%. |
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| Keywords: | Non-heading Chinese cabbage Microsatellite ISSR-PCR Streptavidin-coated magnetic beads adsorption SSR primer |
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