| 新型植物油莎豆DNA提取与SRAP体系优化 |
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| 引用本文： | 赵永国,金梦阳,危文亮.新型植物油莎豆DNA提取与SRAP体系优化[J].中国油料,2011(1):39-43. |
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| 作者姓名： | 赵永国 金梦阳 危文亮 |
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| 作者单位： | 中国农业科学院油料作物研究所,农业部油料作物生物学重点开放实验室,湖北武汉430062 |
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| 基金项目： | 国家自然科学基金（31000727）; 湖北省自然科学基金（2008CDB092）; 农业部油料作物生物学重点开放实验室开放基金（2010-1） |
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| 摘 要： | 以14个不同地理来源的油莎豆品系为实验材料,研究了油莎豆DNA快速提取方法;以SRAP反应体系中DNA模板量、Mg2＋浓度和引物浓度3个因子分别设置3个水平,共配制27个反应体系,对油莎豆SRAP反应体系进行优化。研究结果表明：改良CTAB法可获得较高质量的DNA,参试材料的A260/A280介于1.70～1.98;在27个SRAP反应体系（15μL）中,最优反应体系为DNA模板25ng、Mg2＋1.5mmol/L、引物浓度1μmol/L、dNTPs 0.3mmol/L和Taq酶1U,可扩增出清晰稳定的多态性条带。
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| 关 键 词： | 油莎豆 DNA提取 SRAP 体系优化 |
DNA extraction and establishment of SRAP reaction system in tigernut（Cyperus esculentus） |
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| Authors: | ZHAO Yong-guo JIN Meng-yang WEI Wen-liang |
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| Affiliation: | (Oil Crops Research Institute,CAAS,Key Laboratory of Oil Crops Biology of the Ministry of Agriculture,Wuhan 430062,China) |
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| Abstract: | In this study,fourteen tigernut lines from different geographical areas were used to investigate the effective method of DNA extraction and to construct the optimized SRAP reaction system.Twenty seven reaction systems were designed with three concentration levels of DNA template,magnesium ion and primers.The results showed that the improved method of CTAB contributed to a better extraction of genomic DNA of tigernut,and the A260/280 values of all samples ranged from 1.70 to 1.98.Out of the 27 reaction systems,the optimal system for SRAP analysis had 25ng DNA,1.5mmol/L MgCl2,1μmol/L primers,0.3mmol/L dNTPs and 1U Taq polymerase in 15μL total volume.These results provided technical assistance to evaluate germplasm resource,genetic diversity and molecular breeding in tigernut. |
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| Keywords: | Cyperus esculentus DNA extraction SRAP System optimization |
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