刺山柑总RNA提取方法的比较研究

Comparison of Extraction Methods for Capparis spinosa L.Total RNA

以刺山柑为试材,研究比较了改良SDS提取法、RNApure Plant Plus Reagent法、RNAplant Plus Reagent法和RNAprep Pure Plant Kit法提取总RNA的效果,以期寻找一种适于野生刺山柑叶片高质量总RNA最佳提取方法。结果表明:4种方法提取的总RNA均可见28S和18S2条电泳谱带,RNAprep Pure Plant Kit法获得的RNA图谱条带清晰、明亮、稳定,28S rRNA亮度约是18SrRNA的2倍,OD260/OD280为2.00,产率为175.3μg/g,除RNAprep Pure Plant Kit法能够从叶片中提取到完整性好、纯度高及产率高的总RNA外,其它3种方法提取的总RNA均存在降解及DNA和蛋白等污染,经cDNA-SRAP分析检测,此方法获得的RNA反转录后能扩增出清晰稳定的多态性条带,能够满足后续试验的要求。进一步采用RNAprep Pure Plant Kit法从刺山柑幼苗整株、根、茎及叶中能够获得高质量的总RNA,因此,RNAprep Pure Plant Kit法适合刺山柑总RNA的提取。

With Capparis spinosa L. as material, the method of modified SDS, RNApure Plant Plus Reagent, RNAplant Plus Reagent and RNAprep Pure Plant Kit were compared to find out the optimal method for extracting high quality total RNA from blades in wild Capparis spinosa L.. The results indicated that four kinds of methods to extract total RNA were seen two electrophoretic bands of 28S and 18S,RNAprep Pure Plant Kit method to obtain an RNA bands were clear pattern, bright, stable, brightness of 28S rRNA was two times of 18S rRNA, while the value of OD260/OD280 was 2. 00 and the yield of RNA was 175. 3μg/g,except RNAprep Pure Plant Kit method could be extracted from the leaves to integrity is good, high purity and high yield of total RNA, other three kinds of total RNA extraction are pollution, such as biodegradation and DNA and protein. Further using RNAprep Pure Plant Kit method from Capparis spinosa L seedlings in the whole plant,root, stem and leaf could obtain high quality total RNA, therefore, RNAprep Pure Plant Kit method was suitable for the extraction of total RNA.