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《Journal of Southwest China Normal University(Natural Science)》 2003-01
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Isolation of Total RNA from Dry Seeds of Davidia involucra

LI YunXiang1,2,CHENLi11Dept. of Biology, Sichuan Teachers College, Nanchong 637002, China;2College of Life Science, Sichuan University, Chengdu 610064, China  
After many unsuccessful attempts to obtain biologically active mRNAs from dry seeds of Davidia involucrata using available procedures, an effective procedure modified from Vicient and Delseny (1999) is developed. This method is based on the use of 8 M LiCl in the extraction buffer followed by phenol extractions. The differences with respect to the original method are the inclusions of 2% soluble PVP (MW 10,000, Sigma) and 10 mM cysteine in the extraction buffer to avoid oxidation of polyphenolics, the reduction in the number of phenol extractions, freezing the seeds for 10 minutes instead of using quarts. Our modifications conquer the perishing oxidation of polyphenolics, in their oxidized forms, polyphenols can covalently bind to proteins and nucleic acids; and permit the recovery of an average of 240 μg total RNA per g of the seeds that is suitable for translation in vitro, northern hybridizations, and the construction of cDNA libraries.
【Fund】: 国家自然科学基金资助项目(39770134);; 四川省教育厅重点项目(川教科1999-164)
【CateGory Index】: Q943
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