不同RNA提取方法的效能比较

Comparative Evaluation on Different RNA Extraction Methods

目的比较7种不同的RNA提取方法(6种商业化试剂盒加Trizol有机提取法)的相对提取效率,为法医实践中选择提取高质量的RNA/miRNA提供参考。方法分别使用6种不同的商业化试剂盒以及Trizol有机提取法提取晾干的外周血棉签样本,通过对RNA进行紫外荧光定量检测、琼脂糖凝胶电泳检测和测定样本中内参基因RNU6b的相对含量等方式进行评价。结果各试剂盒所提RNA/miRNA的质量评估和表达分析结果有明显差异,其中Trizol有机提取法和试剂盒RNeasy?Mini可以得到更高质量和纯度的RNA/miRNA, 170μL外周血中所提取的RNA总量可达到(3860.50±182.12)ng、纯度(OD260/280)约为1.84±0.03,且琼脂糖电泳结果显示RNA完整性好,在后续实验中表现良好。结论在所有的提取方法中,综合考虑RNA的数量、质量以及Realtime PCR检测结果,更适合提取外周血RNA的方法为RNeasy?Mini Kit和Trizol有机提取法。

Objective To compare the seven commonly-used commercially-available RNA extraction kits and relevant methods(six commercially available extraction kits plus Trizol organic extraction suite)so as to assess their relative effectiveness of yielding RNA/miRNA.Methods Dried peripheral bloods were taken as the samples for the RNA to extract respectively by the seven selected methods.RNA quality was evaluated with UV spectrophotometry,agar-gel electrophoresis and quantitative PCR expression analysis.Results It was showed that different methods exhibit considerable discrepancy for either the quality or expression levels.Both Trizol organic RNA extraction suite and RNeasy^■Mini Kit can get RNA/miRNA of good quantity and quality(From170μL peripheral blood,the total obtained RNA quantity>1353.50±78.87ng,OD260/280>1.84±0.03),rendering the yielded RNA to be of good integrity as revealed by agar-gel electrophoresis.Conclusion From overall comprehension by the quantity,quality and quantitative PCR-based expression analysis,the RNeasy^■Mini Kit and Trizol are more suitable for extracting RNA/miRNA from peripheral blood.