A Method for Extracting Total RNA of Taxus chinensis at Large-scale

doi:10.13560/j.cnki.biotech.bull.1985.2017-0319

Abstract

Abstract: This study was directed to develop the optimal method for the extraction and purification of total RNA in Taxus chinensis R. First,we investigated the effects of extraction reagent,time,and method on the yield of total RNA from T. chinensis R. using Trizol and CATB extraction methods in respect of RNA concentration and the ratios of OD₂₆₀/OD₂₈₀ and OD₂₆₀/OD₂₃₀. Furthermore,we compared the effects of PVP,β-mercaptoethanol,high salt precipitation,low concentration ethanol precipitation and gel column chromatography on the purification of total RNA. The optimal extraction method was determined as follows：treatment of taxus leaves with Trizol by the action volume of 3：40 in 5 minutes using chloroform and water（1：5）as solvent. It’s also found that column chromatography significantly improved the purity of total RNA of taxus and obtained small molecular RNA with rich miRNA. Finally,the high-quality RNA in concentration of 1000µg/mL with 2.00 - 2.20 of OD₂₆₀/OD₂₈₀ and 1.7 of OD₂₆₀/OD₂₃₀was acquired. Compared with previous methods,this method provides taxus RNA in large-scale with high quality and purity as well as with low cost,which guarantees sufficient materials.

Key words: Taxus chinensis R., total RNA extraction, Trizol method, CATB method, gel column chromatography

CHEN Xiao-feng, ZENG Qi-feng, DENG Xu, ZENG Guang-yao, ZHOU Ying-jun. A Method for Extracting Total RNA of Taxus chinensis at Large-scale[J]. Biotechnology Bulletin, 2017, 33(11): 60-66.

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