葡萄总RNA提取方法的研究An Improved Method for Rapidly Extracting Total RNA from Vitis
张今今,王跃进,王西平,杨克强,杨进孝
摘要(Abstract):
针对葡萄组织中多酚、多糖类物质含量较高的特点,比较了改进SDS/酚法、异硫氰酸胍法和市售总RNA提取试剂盒等不同的提取方法,3种改进方法均能从葡萄幼嫩叶片中提取到总RNA。其中改进的SDS/酚法能有效抑制酚类物质和多糖对总RNA提取的影响,能够从成熟的叶片和完熟果实的果皮中获得质量高、完整性好的总RNA,28SrRNA亮度约为18SrRNA的2倍,A260/A280介于1.8~2.0之间,A260/A230大于2.0,且总RNA产率高,其中幼叶总RNA产率为261.20μg/g,成熟叶产率为191.40μg/g,完熟果皮产率为31.50μg/g,完全适于进行DDRT-PCR、Northernbolt和cDNA文库构建等研究。且该方法具有快速、简单、有效的特点。
关键词(KeyWords): 葡萄;多酚;多糖;SDS/酚法;总RNA提取
基金项目(Foundation): 国家自然科学基金资助项目(39970524和30170653);; 863计划(2001AA241172);; 国家“十五”科技攻关项目(2002BA515B11)。
作者(Author): 张今今,王跃进,王西平,杨克强,杨进孝
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