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

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.