广东药学院学报2017，Vol.33Issue(1)：18-22,5.DOI:10.16809/j.cnki.2096-3653.2016111701
适用于转录组测序的人参根总RNA提取方法的筛选
Screening method of total RNA extraction from Panax ginseng root for RNA-Seq
摘要
Abstract
Objective To screen extracting method for total RNA from Panax ginseng root tissues for RNA-Seq.Methods Panax ginseng grown in Changbai mountain for 15 years was taken as materials. RNAiso Plus Reagent, Trizol, modified CTAB, Total RNA Kit plant extraction reagent method, and RNA prep Pure Plant Kit method were compared and improved to isolate RNA from Panax ginseng root. The total RNA was detected by Agarose gel electrophoresis ( AGE) and Aligent 2100 Bioanalyzer. Results Trizol method and RNAiso Plus method could both obtain high-quality total RNA. 1% Agarose gel electrophoresis showed that 28S and 18S bands were clear and stable, and the values of D (γ) 260 nm/D (γ) 280 nm and D (γ) 260 nm/D (γ) 230 nm were about 2.0. The brightness of 28S with total RNA Trizol method was significantly higher than the 18S, which resulted 2 times in Aligent 2100 Bioanalyzer detection, and the RNA concentration was 485.66 ng/μL. RNA extracted with Prep Pure Plant Kit RNA kit and Total RNA Kit plant were fuzzy, dispersed and degradated. Improved CTAB method could not complete the extraction of total RNA from Panax ginseng root tissues. Conclusion Compared with other methods, the concentration, purity and integrity of total RNA extracted by Trizol method was the best and could meet the requirements of sequencing. It is the preferred method of RNA extraction from Panax ginseng root.关键词
长白山人参/总RNA/多糖多酚/RT-PCR/RNA-SeqKey words
Panax ginseng/total RNA/polyphenolics and polysaccharides/RT-PCR/RNA-Seq分类
医药卫生引用本文复制引用
王颖芳,陈艳琳,王文娟..适用于转录组测序的人参根总RNA提取方法的筛选[J].广东药学院学报,2017,33(1):18-22,5.基金项目
国家自然科学基金青年基金项目(81403195) （81403195）
广东省自然科学基金项目(S2013010015418) （S2013010015418）
