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两种实时定量RT—PCR方法检测miRNAs表达的技术分析
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Journal Article
两种实时定量RT—PCR方法检测miRNAs表达的技术分析
2013
Overview
目的比较两种实时定量反转录聚合酶链反应（RT—qPCR）方法检测microRNAs（miRNAs）表达含量的技术差异，优化不同实验目的和条件下研究miRNAs表达的技术方法。方法取21d和42d两个时间点的SD大鼠关节软骨组织，采用Trizol法提取总RNA备用。选取rnomiR-15b、rno—miR16、rrlo—miR195、rnomiR-497作为研究对象，分别用茎环引物和试剂公司提供的试剂盒方法反转录总RNA，并应用实时定量PCR方法检测这些miRNAs的表达量。提取人血浆中总RNA，用上述两种RT—qPCR方法实时定量检测has—miR16的表达量。结果两种方法检测这些miRNAs表达量，在大鼠21d和42d这两个时间点其表达量变化趋势相同，都呈现增高的趋势，这与我们前期Solexa序结果相同。在血浆中的结果显示，其中茎环引物反转录方法灵敏度相对较高。结论茎环引物法在少量几个重要的miRNAs检测中具有优势，而试剂盒方法适用于大量miRNAs的筛查。
Publisher
西安交通大学医学院遗传学与分子生物学系,陕西西安,710061
Subject
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