从麻疯树胚乳中提取总RNA的快速方法

A Simple Method for Extracting Total RNAs from Endosperm of Jatropha curcas L.

以麻疯树胚乳为材料,建立了一种适合于提取富含油脂、蛋白及多酚、多糖等次生物质的植物总RNA的方法.其中,提取液Ⅰ在1.5%SDS和400mmol/LNaCl盐溶液条件下能快速有效破壁,酸酚/氯仿促进了细胞裂解以及油脂、蛋白的分离.而基于异硫氢酸胍法的提取液Ⅱ进一步强烈抑制RNase的活性,确保RNA不被降解,并以5mol/L高浓度NaCl溶液除去植物组织提取液中的多糖、多酚等次生物质,得到质量较好的总RNA样品.

A method was established for RNA extraction from the endosperm of J. curcas oilseed, which highly containis oil, protein and secondary metabolites, such as polyphenol and polysaccharides. Extraction buffer Ⅰ, which contained SDS and NaCl, could effectively promote cell lysis. And acid phenol-choroform was used to separate DNA and protein. Furthermore, a GITC-based extraction buffer Ⅱ ensured RNase inhibition. High concentration of salt facilitated the separation of nucleic acids from contaminant.