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| 棉花DNA提取及优化SSR反应体系的建立和应用 | |
| 引用本文： | 高文伟,陈全家,曲延英,麻浩,马林,张鹏飞.棉花DNA提取及优化SSR反应体系的建立和应用[J].新疆农业大学学报,2009,32(6):34-37. |
| 作者姓名： | 高文伟 陈全家 曲延英 麻浩 马林 张鹏飞 |
| 作者单位： | 1. 新疆农业大学,农学院/新疆农业大学,生物技术重点实验室,乌鲁木齐,830052;教育部棉花工程研究中心,乌鲁木齐,830052 2. 南京农业大学,南京,210095 3. 新疆农业大学,农学院/新疆农业大学,生物技术重点实验室,乌鲁木齐,830052 |
| 基金项目： | 新疆维吾尔自治区科技厅高技术与发展计划项目，新疆维吾尔自治区高校科研计划 |
| 摘 要： | 对棉花DNA提取程序和SSR反应体系进行了研究。利用正交设计对模板DNA、引物、Taq酶和Mg^2＋浓度4种成分以3个梯度进行优化和选择，结果表明，最优反应体系为15μL，其中含有：10×Taq buffer 1．5μL、Mg^2＋（25mmol／μL）1．5肛L、dNTPs（25mmol／μL）2．0μL、引物（20pmol／μL）各2．0μL、Taq酶（2．0U／μL）0．15μL、DNA模板（25ng）3．0μL、ddH2O补至15μL。采用建立的反应体系，利用8对引物对3个品种进行扩增，6％的变性聚丙烯酰氨凝胶电泳检测结果显示该体系扩增结果清晰稳定。 |
| 关 键 词： | 棉花 正交试验 PCR 反应体系 |
Establishment and Application of Isolation of Genetic DNA and SSR Reaction System in Cotton |
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| GAO Wen-wei,CHEN Quan-jia,QU Yan-ying,MA Hao,MA Lin,ZHANG Peng-fei.Establishment and Application of Isolation of Genetic DNA and SSR Reaction System in Cotton[J].Journal of Xinjiang Agricultural University,2009,32(6):34-37. | |
| Authors: | GAO Wen-wei CHEN Quan-jia QU Yan-ying MA Hao MA Lin ZHANG Peng-fei |
| Institution: | GAO Wen-wei , CHEN Quan-jia, QU Yan-ying, MA Haoa , MA Lin , ZHANG Peng-fei(1. Key Laboratory of Agricultural Biotechnology of Xinjiang,College of Agronomy, Xinjiang Agricultural University, Urumqi 830052, China ; 2. Research Centre for Cotton Engineering, Ministry of Education,Urumqi 830052,Chinas 3. Nanjing Agricultural University,Nanjing 210095,China) |
| Abstract: | In this paper the DNA extraction procedure of cotton and the SSR-PCR reaction system were optimized. Using orthogonal design, four factors including template DNA, primer, Taq polymerase,Mg^2＋ at three grads levels were optimized and selected to establish SSR reaction system for cotton molecular breeding. The results indicated that the optimal reaction system were 15 μL. including 10×buffer 1.5μL, Mg^2＋ （25 mmol/μL）l. 5 μL,dNTPs（25 mmol/μL）2.0 μL,Primer （20 pmol/μL）2.0μL for each one, Taq （2.0 U/μL）0.15 /μL,DNA（25 ng）3.0 /μL,ddH2O 2.85 μL. On optimal conditions,9 pairs of primers were used to analyze 3 eultivars of cotton; PCR products were tested by electrophoresis, using 6% denatured polyacrylamide gels. The results showed this optimal system was stable and reliable. |
| Keywords: | PCR |
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