不同RNA提取方法的效能比较  

Comparative Evaluation on Different RNA Extraction Methods

作　　者：李冉冉 王兵[3] 胡胜[1] 李洋[1] 翟永杰 李彩霞[1,5] 孙启凡[1] 季安全[1] LI Ranran;WANG Bing;HU Sheng;LI Yang;ZHAI Yongjie;LI Caixia;SUN Qifan;JI Anquan(Institute of Forensic Science,Ministry of Public Security&Beijing Engineering Research Center of Crime Scene Evidence Examination,National Engineering Laboratory for Forensic Science,Beijing100038,China;Center of Forensic Science,Jining Medical University,Jining272067,Shandong,China;Weiyang Sub-Bureau of Xi'an Public Security Bureau,Xi'an710016,China;Hohhot Public Security Bureau,Hohhot010090,China;Shanxi Medical University,Taiyuan030001,China)

机构地区：[1]公安部物证鉴定中心北京市现场物证检验工程技术研究中心现场物证溯源技术国家工程实验室,北京100038 [2]济宁医学院司法鉴定中心,山东济宁272067 [3]西安市公安局未央分局,西安710016 [4]呼和浩特市公安局,呼和浩特010090 [5]山西医科大学,太原030001

出　　处：《刑事技术》2018年第6期431-435,共5页Forensic Science and Technology

基　　金："十三五"国家重点研发计划项目(No.2017YFC0803503);公安部科技强警项目(No.2016GABJC17);中央级公益性科研院所基本科研业务费专项资金项目(No.2016JB038;No.2017JB003)

摘　　要：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.

关 键 词：法医遗传学 微RNA 提取方法 定量PCR技术 

分 类 号：DF795.2[医药卫生—法医学]