| 萝卜EST-SSR标记PCR反应体系的建立与优化 |
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| 引用本文： | 付卫民,王淑芬,王秀峰,等.萝卜EST-SSR标记PCR反应体系的建立与优化[J].长江蔬菜,2012(22):6-8. |
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| 作者姓名： | 付卫民 王淑芬 王秀峰 等 |
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| 作者单位： | [1]山东农业大学园艺科学与工程学院,泰安271018 [2]山东省农业科学院蔬菜研究所／山东省设施蔬菜生物学重点实验室／国家蔬菜改良中心山东分中心,泰安271018 |
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| 基金项目： | 山东省现代农业产业技术体系,山东省农业重大应用技术创新课题资助 |
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| 摘 要： | 采用L16(45)正交试验设计优化了萝卜EST-SSR标记的PCR反应体系,建立了适于20μL PCR技术体系的最佳浓度:DNA模板65 ng/20μL,Mg2+0.8 mmol/L,dNTPs 0.25 mmol/L,引物0.8μmol/L,Taq DNA酶0.4 U/20μL,且Mg2+对该体系的影响最大。利用10对EST-SSR引物对该体系的验证结果表明,本研究建立的萝卜EST-SSR PCR体系重复性好、条带清晰、多态性丰富,对萝卜的种质资源的鉴定与评价、分子标记辅助选择育种等多方面都有重要作用。
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| 关 键 词： | 萝卜 EST-SSR PCR 体系优化 |
Establishment and Optimization of EST-SSR Maker PCR System on Radish |
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| FU Weimin,WANG Shufen,WANG Xiufeng,HE Qiwei,LIU Xianxian,HAN Xiaoyu.Establishment and Optimization of EST-SSR Maker PCR System on Radish[J].Journal of Changjiang Vegetables,2012(22):6-8. |
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| Authors: | FU Weimin WANG Shufen WANG Xiufeng HE Qiwei LIU Xianxian HAN Xiaoyu |
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| Institution: | 1.College of Horticulture Science and Engineering,Shandong Agricultural University,Tai＇an 271018; 2.Institute of Vegetable Research,Shandong Academy of Agricultural Sciences/Key Laboratory for Biology of Greenhouse Vegetable of Shandong Province/National Center for Vegetable Improvement,Shandong Branch) |
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| Abstract: | In this paper,we optimized the EST-SSR maker PCR system of radish by using L16（45） orthogonal design.The results showed that the best PCR system in 20 μL was as follows： 65 ng/20 μL DNA template,0.8 mmol/L Mg2＋, 0.25 mmol/L dNTPs,0.8 μmol/L primer,0.4 U/20 μL Taq DNA polymerase,and the greatest factor for this system was the concentration of Mg2＋.The system had been verified by 10 pairs of EST-SSR primers,and the newly established EST-SSR PCR system for radish was fully repeatable with high reliability,clear bands and rich DNA polymorphism,which could be applied in the genetic evaluation of radish germplasm resources and in the molecular assisted selection breeding of radish. |
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| Keywords: | Radish EST-SSR PCR System optimization |
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