| 苜蓿基因组DNA的提取及RAPD反应组成优化探讨 |
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| 引用本文： | 蒿若超,曲春礼,张月学,唐凤兰.苜蓿基因组DNA的提取及RAPD反应组成优化探讨[J].黑龙江农业科学,2006(5):89-92. |
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| 作者姓名： | 蒿若超 曲春礼 张月学 唐凤兰 |
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| 作者单位： | 1. 黑龙江省农科院草业研究所,哈尔滨,150086
2. 宾县农业技术推广中心,宾县,150400 |
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| 摘 要： | 比较了CTAB法和SDS法提取苜蓿基因组总DNA的效果,这两种方法得到的全基因DNA具有较好的完整性,但从DNA的提取纯度可以看出,CTAB法优于SDS法.实验对苜蓿RAPD反应条件进行了优化:在25μL反应体系中,含10×Buffer(20mM MgCl2),1UTaq酶,25ng的模板DNA,150 mol/L的dNTPs,0.2 mol/L引物.PCR反应程序为:94℃预变性3 min;94℃变性15 s,37℃退火30 s,72℃延伸1 min,35个循环;72℃延伸10 min;最后4℃保存.
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| 关 键 词： | 苜蓿 基因组DNA提取 RAPD反应 |
| 文章编号： | 1002-2767（2006）05-0089-04 |
| 收稿时间： | 2006-04-05 |
| 修稿时间： | 2006年4月5日 |
Study on the Methods of Extraction and Purification for Alfalfa DNA and the Establishment and Optimization for RAPD Reaction System |
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| HAO Ruo-chao,QU Chun-li,ZHANG Yue-xue,TANG Fen-lan.Study on the Methods of Extraction and Purification for Alfalfa DNA and the Establishment and Optimization for RAPD Reaction System[J].Heilongjiang Agricultural Science,2006(5):89-92. |
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| Authors: | HAO Ruo-chao QU Chun-li ZHANG Yue-xue TANG Fen-lan |
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| Abstract: | Two methods(SDS method and CTAB method) for DNA extracting were compared,and whole DNA could be got by two methods.But took purity of DNA into account,CTAB method was better than SDS method.Meantime,PCR amplified procedure was established and PCR amplified conditions were defined on the factors of Taq DNA polymerase,DNA density,dNTPs density,primer density,and pre-denaturating temperature. |
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| Keywords: | alfalfa DNA extraction RAPD reaction |
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