在提取缓冲液中加入皂土有效地去除了蛋白质并抑制RNAse,建立了一种高效的植物RNA提取方法。以麻疯树幼叶为材料,分别用TRIZOL,异硫氰酸胍法,SDS-KAc法和新创皂土法提取总RNA进行比较和验证,琼脂糖凝胶电泳和紫外光谱分析结果表明,只有皂土法能提出质量高,完整性好的RNA。进一步以皂土法提取的18SrRNA为模板的RT-PCR结果分析表明,用该法提取的RNA能用于分子克隆与基因表达分析等后继分子生物学实验。该方法简便,快速,不失为一种经济高效的RNA提取方法。
A new and efficient method for isolation of plant RNAs was developed by adding bentonite into extraction buffer in order to get rid of protein and restrain RNAse. The electrophoretic patterns of nucleic acids and absorbance at 230 nm, 260 nm and 280 nm in a UV-Vis spectrophotometer revealed the extraction with this method can obtain RNAs with good integrity and purity without any apparent DNA contamination from the plant materials rich in with polysaccharide and polyphenol like Jatropha curcas leaves, to which TRIZOL reagent, SDS-KAc solution and Guanidine isothiocyanate solution failed. Furthermore, the result of nuclear gene ( 18S rRNA gene) amplified by RT-PCR indicated that the RNAs prepared with this method can meet the needs of most molecular biological experiments including gone cloning and expression analysis.