摘要
利用正交试验法优化血清miRNA的提取方法,为进一步研究血清miRNA作为生物标记物提供方法学基础.本文以hsa-miRNA-223-3p,hsa-miRNA-483-5p和hsa-miRNA-16为检测目标,通过比较Trizol法和Trizol+酚/氯仿法的miRNA提取效率和qPCR检测结果,应用正交试验选择出最优实验方案.结果表明血清miRNA的最优提取方法为:血清用DEPC水4倍稀释,离心条件为4℃,12 000r·min-1,离心时间为10min,Trizol法提取.
To provide the methodological basis for serum miRNA as a biomarker,an orthogonal array design for optimizing miRNA extraction from serum was studied.In this study,RNA extraction efficiency and expression levels of hsa-miRNA-223-3p,hsa-miRNA-483-5pand hsa-miRNA-16 were measured by real-time PCR to make a comparison between single Trizol method,and combination of Trizol and phenol/chloroform method.The results showed that the optimum miRNA extraction method was that serum was diluted 4times with DEPC water,centrifuged at 12 000r·min-1 for 10 min at 4℃ and then used to extract miRNA by Trizol method.
出处
《河南师范大学学报（自然科学版）》
CAS
北大核心
2015年第4期100-103,共4页
Journal of Henan Normal University(Natural Science Edition)
基金
国家自然科学基金(31301883)
新乡市重点科技攻关项目(ZG13002)
