Expression profiling of microRNA using real-time quantitative PCR, how to use it and what is available

被引：268

作者：

Benes, Vladimir ^{[2
]}

Castoldi, Mirco ^{[1
,3
]}

机构：

[1] Heidelberg Univ, Dept Pediat Hematol Oncol & Immunol, D-69120 Heidelberg, Germany

[2] European Mol Biol Lab, D-69117 Heidelberg, Germany

[3] Heidelberg Univ, Mol Med Partnership Unit, D-69120 Heidelberg, Germany

来源：

METHODS | 2010年 / 50卷 / 04期

关键词：

microRNA; qPCR; GENE-EXPRESSION; RT-PCR; MESSENGER-RNA; QUANTIFICATION; LNA; CANCER; PROBES; METABOLISM; MICROARRAY; SOFTWARE;

D O I：

10.1016/j.ymeth.2010.01.026

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

We review different methodologies to estimate the expression levels of microRNAs (miRNAs) using real-time quantitative PCR (qPCR). As miRNA analysis is a fast changing research field, we have introduced novel technological approaches and compared them to existing qPCR profiling methodologies. qPCR also remains the method of choice for validating results obtained from whole-genome screening (e.g. with microarray). In contrast to presenting a stepwise description of different platforms, we discuss expression profiling of mature miRNAs by qPCR in four sequential sections: (1) cDNA synthesis; (2) primer design; (3) detection of amplified products; and (4) data normalization. We address technical challenges associated with each of these and outline possible solutions. (C) 2010 Elsevier Inc. All rights reserved.

引用

页码：244 / 249

页数：6

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