美洲南瓜种皮RNA提取方法的比较

Comparison of Different Extraction Methods of Total RNA from Seed Coat of Cucurbita pepo

为探索提取高质量RNA的方法,采用改良CTAB法、SDS法、异硫酸氰胍法和Trizol法从美洲南瓜种皮中提取总RNA。结果表明:改良CTAB法提取的RNA中28 S rRNA和18 S rRNA 2条带清晰,且28 S rRNA在亮度约为18 S rRNA的2倍,两条带之间无弥散现象;OD260/OD280值为1.996 5,OD260/OD230为2.235 7,RNA产率为161μg/g。异硫酸氰胍法提取的总RNA纯度较高,但产量较低;SDS法和Trizol法提取的总RNA产量较低,且纯度不高。改良的CTAB法从种皮中提取的RNA质量好、产率高、完整性强,是从富含次生代谢物质的种皮中提取高质量总RNA的有效方法。

The improved CTAB,SDS,guanidinium isothiocyanate and trizol extraction methods were tested in total RNA extraction from seed coat of Cucurbita pepo to compare the extraction effect for selecting the efficient method.The results showed that the optimum extraction method was the improved CTAB method with good quality RNA,high productivity and strong integrity,the brands of 28 S and 18 S of total RNA isolated by the CTAB method both were clear,the brightness of 28 S rRNA was about 2 times that of 18S rRNA,there was no diffusing phenomenon between two bands,the values of OD260/OD280 and OD260/OD230 were 1.996 5 and 2.235 7,and RNA productivity was 161 μg/g.