一种适宜棉花microRNA实时定量分析的总RNA提取方法  

A Protocol for Total RNA Isolation Suitable for Real-time PCR Analysis of Cotton MicroRNAs

作　　者：余道乾[1] WANG Gaskin 杜雄明[1] 

机构地区：[1]中国农业科学院棉花研究所/棉花生物学国家重点实验室.河南安阳455000

出　　处：《棉花学报》2013年第5期467-470,共4页Cotton Science

基　　金：国家973项目(2004CB117306)

摘　　要：为满足棉花组织micro RNA结构与功能研究的需要,建立了棉花高质量的总RNA提取方法。以陆地棉纤维和胚珠为供试材料,采用改良的异硫氰酸胍-硫氰酸铵-酸酚法提取总RNA,利用stem-loop RT-qPCR法鉴定成熟的miRNA。结果表明此方法能得到高质量的总RNA,并且miRNA的回收率较高,足以满足microRNA水平的实时定量分析。A method for isolation of total RNA species is presented. The modified guanidinium isothiocyanate- ammonium thio- cyanate-acid phenol method was used to extract total RNA from fiber and ovule of cotton （Gossypium hirsutum L.）. The stem-loop Real-time PCR method was selected to identify mature miRNA. Experiments validated that the method we used was suitable for quantitative analysis ofmiRNA because of the high quality of total RNA and high recycling efficiency of miRNA.

关 键 词：棉花 总RNA 提取 MICRORNA 实时定量 

分 类 号：S562[农业科学—作物学]