An optimized method to obtain high-quality RNA from cassava storage root
An optimized method to obtain high-quality RNA from cassava storage root
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从木薯贮藏根中获取高质量RNA的优化方法
DOI：
10.1007/s13205-019-1608-9
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发表时间：
2019-04-01
期刊：
影响因子：
2.8
通讯作者：
Wang, Zhen-Yu
中科院分区：
文献类型：
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作者：
Guan, Lulu;Ma, Xiaowen;Wang, Zhen-Yu
高质量、完整的RNA提取是真核生物分子生物学研究的关键。然而，木薯贮藏根中含有大量的多糖和多酚类化合物，因此从木薯贮藏根中提取RNA引起了人们的极大关注。本研究比较了4种RNA提取方法对木薯块根RNA提取的影响。结果表明，改良TM法（MTM）是一种省时、低成本、高质量、高产量的RNA提取方法。利用改进的方法对从贮藏根总RNA中筛选出的4个基因进行qPCR分析。改进的方案产生4.18-5.94 μg RNA/g鲜重。RNA样品的A260/280比值范围为2.14至2.17。RIN值范围为7.2至8.0。重要的是，通过MTM分离的总RNA成功地用于文库构建和转录组测序。因此，我们提供了一种高效，低成本的方法，MTM，提取高质量和数量的RNA从木薯储藏根。
Extracting RNA with high quality and integrity is crucial for molecular biology studies in eukaryotes. However, RNA isolation from cassava storage root raises a great concern because it contains large amounts of polysaccharides and polyphenol compounds. In the current study, four RNA extraction methods were evaluated for extracting RNA from cassava storage root. We found that the modified TM method (MTM) is timesaving and low-cost extraction method with high quality and quantities of RNA. The effectiveness of the improved method was assessed for qPCR analysis of four selected genes from total RNA of storage root. The improved protocol generated 4.18–5.94 µg RNA/g fresh weight. An A260/280 ratios of RNA samples are ranged from 2.14 to 2.17. The RIN values are ranged from 7.2 to 8.0. Importantly, isolated total RNA by MTM was successfully used for library construction and transcriptome sequencing. Therefore, we provide an efficient and low-cost method, MTM, for extracting high quality and quantities of RNA from cassava storage root.