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| 葡萄基因组DNA的提取及RAPD反应体系的优化 | |
| 引用本文： | 王晶晶,吴森林,张付春,高疆生.葡萄基因组DNA的提取及RAPD反应体系的优化[J].新疆农业科学,2010,47(6):1066-1070. |
| 作者姓名： | 王晶晶 吴森林 张付春 高疆生 |
| 作者单位： | 1. 塔里木大学植物科学学院,新疆阿拉尔,843300;新疆生产建设兵团塔里木盆地生物资源保护利用重点实验室,新疆阿拉尔,843300 2. 莎车县农业技术推广中心,新疆喀什,844700 3. 新疆农业科学院园艺作物研究所,乌鲁木齐,830091 4. 新疆生产建设兵团塔里木盆地生物资源保护利用重点实验室,新疆阿拉尔,843300 |
| 基金项目： | 国家"十一五"科技支撑计划 |
| 摘 要： | 【目的】建立适合该地区部分葡萄品种RAPD扩增反应体系。【方法】采用阿克苏地区部分葡萄品种(系)为试材,采用改良的CTAB法提取葡萄基因组DNA,对影响葡萄RAPD反应的Mg2+、Taq酶、dNTPs、引物、模板DNA浓度进行优化。【结果】其最佳反应体系为:模板DNA 100 ng,dNTPs 0.1 mmol/L,Mg2+2.0 mmol/L,引物0.3μmol/L,Taq酶1.5 U,反应体系总体积为20μL。【结论】改良的CTAB法具有步骤少、费用低、效率高等特点,比较适用于葡萄基因组DNA的提取。优化后的体系,扩增产物稳定,条带清晰,重复性好。 |
| 关 键 词： | 葡萄 DNA提取方法 RAPD 优化 |
Grape Genomic DNA Extraction and Optimization of RAPD Reaction System |
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| WANG Jing-jing,WU Sen-lin,ZHANG Fu-chun,GAO Jiang-sheng.Grape Genomic DNA Extraction and Optimization of RAPD Reaction System[J].Xinjiang Agricultural Sciences,2010,47(6):1066-1070. | |
| Authors: | WANG Jing-jing WU Sen-lin ZHANG Fu-chun GAO Jiang-sheng |
| Institution: | WANG Jing-jing1,2,WU Sen-lin3,ZHANG Fu-chun4,GAO Jiangsheng2(1.College of Plant Science and Technology,Tarim University,Alar Xinjiang 843300,China,2.Key Laboratory of Protection & Utilization of Biological Resources in Tarim Basin,Xinjiang Production and Construction Corps,3.Agricultural Technology Extension Centre of Shache County,Kashi Xinjiang 844700,4.Institute of Horticultureal Crops,Xinjiang Academy of Agricultural Sciences,Urumqi 830091,China) |
| Abstract: | 【Objective】To establish the adaptive RAPD amplification reaction system .【Method】Several grape varieties in Akesu were chosen as test materials,genomic DNA was extracted by improved CTAB,Mg2+,Taq polymeras,dNTPs,DNA template concentration and primer affecting RAPD reactions were optimized.【Result】 The best reaction system was DNA template 100 ng,dNTPs 0.1 mmol/L,Mg2+ 2.0 mmol/L,primer 0.3 umol/L,Taq polymeras 1.5 U in 20 μL reaction system.【Conclusion】The improved CTAB method has the advantages of reduced steps,low-cost and high-efficiency,which is more suitable for grape genomic DNA extraction.For the optimized system,the amplification products were stable,bands were clear and repeatability was good. |
| Keywords: | RAPD |
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