The Improvement on the Protocols of Total RNA Extraction Kits for RNA-Seq from Grape Leaves

Abstract

Abstract: Commercial RNA extraction kits were wildly used in obtaining high quality RNA samples from plants. To seek the optimal method for the extraction of high quality RNA in RNA Expression Sequences Array(RNA-Seq), the topic of present article is to evaluate and optimize the effects of three common RNA extraction kits on leaves samples from grapevine(Vitis vinifera cv. Red Globe). The quantities of RNA were analyzed between the recommendatory and optimal usages of three kits in term of times. Results showed that the optimized procedure for RNA extraction displayed higher yield and better integrity on total RNA. There are a ratios of OD₂₆₀/OD₂₈₀ from 1.8 to 2.2 and further OD₂₆₀/OD₂₃₀ratio no less than 2, resulted in a integrate 28S and 18S bands by electrophoresis. The extracted samples also kept their quality and gene coverage from RNA-Seq sequencing. Our research demonstrated that the prolonged incubation time for samples preparation and other procedures would significantly benefit the separation of total RNA from grape leaves(Vitis vinifera cv. Red Globe)with a higher purity and integrity.

Key words: Vitis vinifera cv., Red Globe, RNA extaction, Kit assay, RNA-Seq

Yang Xiaoyan, Zhang Bo, Huang Fang'ai, Yan Huan, Li Yuerong, Zheng Qiusheng. The Improvement on the Protocols of Total RNA Extraction Kits for RNA-Seq from Grape Leaves[J]. Biotechnology Bulletin, 2013, 0(6): 215-220.

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