| 一种适合葡萄多种组织的总RNA提取方法 |
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| 引用本文： | 李红熙,徐美隆,杨智.一种适合葡萄多种组织的总RNA提取方法[J].中国农学通报,2012,28(7):155-159. |
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| 作者姓名： | 李红熙 徐美隆 杨智 |
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| 作者单位： | 1. 国家经济林木种苗快繁工程技术研究中心,银川750004;宁夏大学生命科学学院,银川750021
2. 国家经济林木种苗快繁工程技术研究中心,银川750004;种苗生物工程国家重点实验室,银川750004 |
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| 基金项目： | 宁夏回族自治区科技攻关计划项目（国际合作专项）“宁夏葡萄脱毒与病毒检测技术体系研究” 宁夏回族自治区科技攻关计划重大专项“宁夏贺兰山东麓葡萄酒产业关键技术研究与示范” |
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| 摘 要： | 为了解决葡萄组织中总RNA提取难度大的问题，以感染葡萄卷叶病毒-3的葡萄品种‘蛇龙珠’为试验材料，采用CTAB结合SDS法对总RNA提取方法进行了研究，建立了一套适合葡萄叶片、叶柄及韧皮部等多种组织总RNA提取的方法。结果表明，该方法在多种组织中均能获得高质量的RNA，OD260/OD280=1.8~1.9，电泳结果表明，所得RNA完整性好，无降解。同时用所得RNA进行了GLRaV-3的检测，电泳结果表明该法所得RNA可满足葡萄病毒病RT-PCR检测的质量要求。该方法操作简单，结果稳定，对其他富含多酚、多糖类植物组织总RNA的提取具有借鉴意义。
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| 关 键 词： | 油茶 油茶 硬脂酰-酰基载体蛋白脱饱和酶 原核表达载体构建 |
| 收稿时间： | 2011/11/15 0:00:00 |
| 修稿时间： | 2012/1/13 0:00:00 |
A Method Suited to Extract Total RNA from Various Tissues of Grapevine |
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| Li Hongxi
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Xu Meilong
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Yang Zhi.A Method Suited to Extract Total RNA from Various Tissues of Grapevine[J].Chinese Agricultural Science Bulletin,2012,28(7):155-159. |
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| Authors: | Li Hongxi Xu Meilong Yang Zhi |
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| Institution: | 1 The National Center of Research and Engineering Technology of Economic Forest Tree Speedy Propagation , Yinchuan 750004; 2 College of Life Science, Ningxia University , Yinchuan 750021; 3 State Key Laboratory of Seeding Bio-engineering ,Yinchuan 750004) |
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| Abstract: | In order to solve the problem of extracting total RNA from grape tissues, the author took the leaf-roll associated virus 3 infected grapes varieties ＇ Cabernet Gernischet ＇ as materials, the method of CTAB combined with SDS was studied in this paper, a suitable method for total RNA extraction from leaves, petioles and phloem of grapevine was established. The results indicated that high-quality RNA could be obtained by this method, with OD 260 /OD 280 =1.8-1.9, without degradation. The results showed that the RNA obtained could be used for RT-PCR to detect grapevine viruses. The method was simple, the results were stable. It also gave better suggestions to extract total RNA of tissues which were rich in polyphenol and polysaccharides. |
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| Keywords: | grape extraction of total RNA virus detection |
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