qPCR-based methods for expression analysis of miRNAs

被引：104

作者：

Forero, Diego A. ^{[1
,2
]}

Gonzalez-Giraido, Yeimy ^{[3
]}

Castro-Veges, Luis J. ^{[4
,5
]}

Barreto, George E. ^{[3
]}

机构：

[1] Univ Antonio Narino, Sch Med, Lab NeuroPsychiat Genet, Biomed Sci Res Grp, Bogota, Colombia

[2] Univ Antonio Narino, Sch Med, PhD Program Hlth Sci, Bogota, Colombia

[3] Pontificia Univ Javeriana, Dept Nutr & Bioquim, Bogota, Colombia

[4] INSERM, UMR970, Paris Cardiovasc Res Ctr, Equipe Labellisee Ligue Canc, Paris, France

[5] Univ Paris 05, Sorbonne Paris Cite, Fac Med, Paris, France

来源：

BIOTECHNIQUES | 2019年 / 67卷 / 04期

关键词：

expression analysis; microRNAs; molecular assays; non-coding RNAs; real-time PCR; REAL-TIME PCR; POLYMERASE-CHAIN-REACTION; RT-PCR; MICRORNA EXPRESSION; GENE-EXPRESSION; DIGITAL PCR; QUANTIFICATION; NORMALIZATION; ASSAY; IDENTIFICATION;

D O I：

10.2144/btn-2019-0065

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Several approaches for miRNA expression analysis have been developed in recent years. In this article, we provide an updated and comprehensive review of available qPCR-based methods for miRNA expression analysis and discuss their advantages and disadvantages. Existing techniques involve the use of stem-loop reverse transcription-PCR, polyadenylation of RNAs, ligation of adapters or RT with complex primers, using universal or miRNA-specific qPCR primers and/or probes. Many of these methods are oriented towards the expression analysis of mature miRNAs and few are designed for the study of pre-miRNAs and pri-miRNAs. We also discuss findings from articles that compare results from existing methods. Finally, we suggest key points for the improvement of available techniques and for the future development of additional methods.

引用

页码：192 / 199

页数：8

共 89 条

[1] Two-tailed RT-qPCR: a novel method for highly accurate miRNA quantification [J].

Androvic, Peter ;

Valihrach, Lukas ;

Elling, Julie ;

Sjoback, Robert ;

Kubista, Mikael .

NUCLEIC ACIDS RESEARCH, 2017, 45 (15)

[2]

[Anonymous], J VIS EXP

[3]

[Anonymous], BIOCH BIOPHYS ACTA G

[4] Specific and sensitive quantitative RT-PCR of miRNAs with DNA primers [J].

Balcells, Ingrid ;

Cirera, Susanna ;

Busk, Peter K. .

BMC BIOTECHNOLOGY, 2011, 11

[5] Identification of cytokine-induced modulation of microRNA expression and secretion as measured by a novel microRNA specific qPCR assay [J].

Benes, Vladimir ;

Collier, Paul ;

Kordes, Claus ;

Stolte, Jens ;

Rausch, Tobias ;

Muckentaler, Martina U. ;

Haeussinger, Dieter ;

Castoldi, Mirco .

SCIENTIFIC REPORTS, 2015, 5

[6] A tool for design of primers for microRNA-specific quantitative RT-qPCR [J].

Busk, Peter K. .

BMC BIOINFORMATICS, 2014, 15

[7] Variability of the Reverse Transcription Step: Practical Implications [J].

Bustin, Stephen ;

Dhillon, Harvinder S. ;

Kirvell, Sara ;

Greenwood, Christina ;

Parker, Michael ;

Shipley, Gregory L. ;

Nolan, Tania .

CLINICAL CHEMISTRY, 2015, 61 (01) :202-212

[8] The MIQE Guidelines: Minimum Information for Publication of Quantitative Real-Time PCR Experiments [J].

Bustin, Stephen A. ;

Benes, Vladimir ;

Garson, Jeremy A. ;

Hellemans, Jan ;

Huggett, Jim ;

Kubista, Mikael ;

Mueller, Reinhold ;

Nolan, Tania ;

Pfaffl, Michael W. ;

Shipley, Gregory L. ;

Vandesompele, Jo ;

Wittwer, Carl T. .

CLINICAL CHEMISTRY, 2009, 55 (04) :611-622

[9] The UCSC Genome Browser database: 2018 update [J].

Casper, Jonathan ;

Zweig, Ann S. ;

Villarreal, Chris ;

Tyner, Cath ;

Speir, Matthew L. ;

Rosenbloom, Kate R. ;

Raney, Brian J. ;

Lee, Christopher M. ;

Lee, Brian T. ;

Karolchik, Donna ;

Hinrichs, Angie S. ;

Haeussler, Maximilian ;

Guruvadoo, Luvina ;

Gonzalez, Jairo Navarro ;

Gibson, David ;

Fiddes, Ian T. ;

Eisenhart, Christopher ;

Diekhans, Mark ;

Clawson, Hiram ;

Barber, Galt P. ;

Armstrong, Joel ;

Haussler, David ;

Kuhn, Robert M. ;

Kent, W. James .

NUCLEIC ACIDS RESEARCH, 2018, 46 (D1) :D762-D769

[10] Telomere length measurement by a novel monochrome multiplex quantitative PCR method [J].

Cawthon, Richard M. .

NUCLEIC ACIDS RESEARCH, 2009, 37 (03)

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