阿魏侧耳菌丝体RNA提取方法的比较与研究

采用一步法、异硫氰酸胍—巯基乙醇联合变性法和STE法3种方法,分别对阿魏侧耳总RNA的提取效果进行比较。结果表明,一步法难以提取RNA,异硫氰酸胍—巯基乙醇联合变性法提取RNA效果不理想,存在蛋白污染,这2种方法不适于富含多糖的阿魏侧耳总RNA的提取；STE法提取阿魏侧耳总RNA质量高、完整性好、成功率高,经琼脂糖凝胶电泳、紫外分光光度计检测,提取的总RNA具有清晰的28S,18S,5S三条带,且28S的亮度是18S的2倍左右,OD260/OD280比值为18～20,可作为阿魏侧耳总RNA提取的首选方法。用此方法来提取阿魏侧耳液体发酵不同时期的RNA,可以满足进一步分子生物学研究的要求。

Comparison of the RNA isolation methods for Pleurotus ferulae Lenzi hyphostroma

Total RNA of Pleurotus ferulae Lenzi was extracted by single-step method, STE method and guanidinium isothiocyanate method, whose isolation effects were compared. The results showed that it was difficult to extract total RNA from Pleurotus ferulae Lenzi by single\|step method. Total RNA could be extracted by guanidinium isothiocyanate method, but RNA extracted was immingled by some protein and was not enough for molecular biology experiment. Single\|step method and guanidiniu isothiocyanate method were not suitable for RNA extraction from Pleurotus ferulae Lenzi because of its high amylase content. Total RNA extracted by STE method had high quality, which is the best method of total RNA isolation from Pleurotus ferulae Lenzi. The quality of total RNA was analyzed with agarose gel electrophoresis and nucleic acid and ultraviolet spectrophotometer. Three bright bands were observed in agarose gel electrophoresis as 28S, 18S and 5S. The band of 28S was twice wider than that of 18S, and the value of OD260/OD280 was 18 to 20.These results demonstrated that the quality and purity of RNA obtained by the method of STE could meet the demands of molecular biology experiment．