Optimization of routine trizol extraction method for promoting the production of serum microRNA

VernacularTitle:常规TRIzol法提取血清microRNA的条件改进
Author: Changxuan CHEN ; Lei WANG ; Wei ZHAO ; Fengzhi SHAN ; Xin WANG ; Xiaochao GAO
Publication Type:Journal Article
Keywords: MicroRNAs/BL/IP; Polymerase chain reaction
From: Journal of Chinese Physician 2015;17(8):1204-1206
CountryChina
Language:Chinese
Abstract: Objective To investigate which methods can promote the serum microRNA extract production.Methods The sera of healthy persons were treated with 10％ SDS (sodium dodecyl sulfate) and/or sonication at first,then extracted miRNA by routine Trizol method,or the serum miRNA was just extracted by routine Trizol method.The contents of serum miRNA-218 and miRNA-346 were detected by quantitative reverse transcription polymerase chain reaction (qRT-PCR) technique,and qRT-PCR relative quantities among different groups were compared to evaluate influence of different methods on microRNA extract efficacy.Results Compared to routine Trizol extraction method,the qRT-PCR relative quantities of miRNA-218 and miRNA-346 were improved when serum was treated with 10％ SDS and/or sonication at first (P ＜0.05),which was most obviously improved when treated with 10％ SDS combined with sonieation.Conclusions The method that serum was treated with SDS incubation and sonication at first can promote sensitivity of serum microRNA detection.