文献类型：专利

英文题名：Performing PCR analysis for detecting target microRNA, involves stabilizing combination of forward primer with template, initiating PCR reaction, combining dye, determining fluorescence signal intensity and calculating target microRNA

作者：YE B;YU C;YIN B

机构：[1]UNIV EAST CHINA SCI & TECHNOLOGY

申请号：CN103320519-A

申请日：2013-07-12

公开日：2013-09-25

语种：英文

收录：DERWENT

摘要：NOVELTY - Method for performing PCR analysis to detect microRNA, involves stabilizing the combination of a forward primer with a DNA amplification template, extending the forward primer, initiating a PCR reaction under the combined action of the extended forward primer and a reverse primer to obtain double-stranded DNA, combining a dye SYBR (RTM: cyanine dye) Green I with the double-stranded DNA, determining the fluorescence signal intensity in a reaction system in real time, comparing with a standard work curve, and calculating to obtain the concentration of the target microRNA. USE - The method is useful for performing PCR analysis for the quantitative detection of microRNA based on a base stacking hybridization principle, where the microRNA is let-7a, miR-141, miR-21 and miR-200b (claimed) in the biological samples of tissues, blood or cells. ADVANTAGE - The method exhibits high sensitivity and strong specificity, and is simple and cost-effective to perform. DETAILED DESCRIPTION - Method for performing PCR analysis for the quantitative detection of microRNA based on a base stacking hybridization principle, involves stabilizing the combination of a forward primer with a DNA amplification template through a base stacking hybridization effect after the combination of target microRNA with the DNA amplification template, extending the forward primer under the action of a DNA polymerase, initiating a PCR reaction under the combined action of the extended forward primer and a reverse primer to obtain double-stranded DNA, combining a dye SYBR (RTM: cyanine dye) Green I with the double-stranded DNA, determining the fluorescence signal intensity in a reaction system in real time, comparing with a standard work curve, and calculating to obtain the concentration of the target microRNA.