葡萄基因组DNA的提取及RAPD反应体系的优化  

Grape Genomic DNA Extraction and Optimization of RAPD Reaction System

作　　者：王晶晶[1,2] 吴森林[3] 张付春[4] 高疆生[2] 

机构地区：[1]塔里木大学植物科学学院,新疆阿拉尔843300 [2]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 [3]莎车县农业技术推广中心,新疆喀什844700 [4]新疆农业科学院园艺作物研究所,乌鲁木齐830091

出　　处：《新疆农业科学》2010年第6期1066-1070,共5页Xinjiang Agricultural Sciences

基　　金：国家"十一五"科技支撑计划"葡萄品种优选及优质高效栽培技术研发与示范"(2007BAD36B08)

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。[ 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, Mg^2＋, 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. I mmol/L, Mg^2＋ 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, hands were clear and repeatability was good.

关 键 词：葡萄 DNA提取方法 RAPD 优化 

分 类 号：S663.1[农业科学—果树学] S188[农业科学—园艺学]