水稻成熟种子总RNA提取方法的比较研究  

Comparative Study on the Methods for Extracting Total RNA from Mature Rice Seeds

作　　者：廉添添 袁会峰 孙强 许园园 刘慧 LIAN Tiantian;YUAN Huifeng;SUN Qiang;XU Yuanyuan;LIU Hui(School of Life Science and Technology,Sanquan College,Xinxiang Medical University,Xinxiang,Henan 453003,China)

机构地区：[1]新乡医学院三全学院生命科学技术学院,河南新乡453003

出　　处：《杂交水稻》2024年第4期36-41,共6页Hybrid Rice

基　　金：河南省科技攻关计划项目(222102310436);河南省高等学校重点科研项目(21B310008);新乡医学院三全学院科技创新团队(STD201704)。

摘　　要：采用PLANTeasy植物RNA提取试剂盒法(PLANTeasy法)、Trizol法、EASYspin植物RNA快速提取试剂盒法(EASYspin法)、CTAB(十六烷基三甲基溴化铵)-LiCl沉淀法、改良CTAB+过柱法等5种方法提取水稻成熟种子的总RNA,通过琼脂糖凝胶电泳、紫外分析和安捷伦2100生物分析仪对总RNA样品进行质量与纯度检测,并以反转录PCR(RT-PCR)检测actin基因的目的条带,旨在明确获得水稻成熟种子高质量总RNA的最佳提取方法。结果显示,EASYspin法和改良CTAB+过柱法提取的总RNA样品的质量与纯度较高,以其提取的总RNA为模板并通过RT-PCR扩增均可获得明亮、单一的目的基因条带。尤其是改良CTAB+过柱法提取的总RNA产率高且完整性好,更适合水稻种子总RNA的提取。In order to find out the optimal extraction method for obtaining high-quality total RNA from mature rice seeds,five extraction methods including PLANTeasy plant RNA extraction kit(PLANTeasy method),Trizol method,EASYspin plant RNA rapid extraction kit(EASYspin method),CTAB(hexadecyltrimethy ammonium bromide)-LiCI precipitation method and modified CTAB+column method were studied comparatively.The quality and purity of total RNA samples were detected by agarose gel electrophoresis,ultraviolet analysis and Agilent 2100 biological analyzer,and the target band of actin gene was detected by reverse transcription PCR(RT-PCR).The results showed that the quality and purity of total RNA samples extracted by the EASYspin method and the modified CTAB+column method were relatively higher than those by the other three methods.A bright and single band of the target gene was obtained by RT-PCR amplification using total RNA extracted by these two methods as template.Particularly,the modified CTAB+column method can obtain a high yield and good integrity of total RNA,so it is more suitable for the extraction oftotal RNA fromrice seeds.

关 键 词：水稻种子 RNA提取 改良CTAB+过柱法 

分 类 号：S511.01[农业科学—作物学] S311