A practical approach to RT-qPCR-Publishing data that conform to the MIQE guidelines

被引：545

作者：

Taylor, Sean ^{[1
]}

Wakem, Michael ^{[1
]}

Dijkman, Greg ^{[1
]}

Alsarraj, Marwan ^{[1
]}

Nguyen, Marie ^{[1
]}

机构：

[1] Biorad Labs Inc, Hercules, CA 94547 USA

来源：

METHODS | 2010年 / 50卷 / 04期

关键词：

RELATIVE QUANTIFICATION; PCR ANALYSIS;

D O I：

10.1016/j.ymeth.2010.01.005

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Given the highly dynamic nature of mRNA transcription and the potential variables introduced in sample handling and in the downstream processing steps (Carson et al. (2009) [4]), a standardized approach to each step of the RT-qPCR workflow is critical for reliable and reproducible results. The MIQE provides this approach with a checklist that contains 85 parameters to assure quality results that will meet the acceptance criteria of any journal (Bustin et al. (2009) [1]). In this paper we demonstrate how to apply the MIQE guidelines (www.rdml.org/miqe) to establish a solid experimental approach. (C) 2010 Published by Elsevier Inc.

引用

收藏

页码：S1 / S5

页数：5

相关论文

共 14 条

[1] 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

[2]

Denisov V, 2008, BIORAD B

[3]

Fleige Simone, 2006, Molecular Aspects of Medicine, V27, P126, DOI 10.1016/j.mam.2005.12.003

[4] Unreliable Real-Time PCR Analysis of Human Endogenous Retrovirus-W (HERV-W) RNA Expression and DNA Copy Number in Multiple Sclerosis [J].

Garson, Jeremy A. ;

Huggett, Jim F. ;

Bustin, Stephen A. ;

Pfaffl, Michael W. ;

Benes, Vladimir ;

Vandesompele, Jo ;

Shipley, Gregory L. .

AIDS RESEARCH AND HUMAN RETROVIRUSES, 2009, 25 (03) :377-378

[5]

GINGRICH J, 2006, BIORAD B, V5452

[6] The lack of a systematic validation of reference genes:: a serious pitfall undervalued in reverse transcription-polymerase chain reaction (RT-PCR) analysis in plants [J].

Gutierrez, Laurent ;

Mauriat, Melanie ;

Guenin, Stephanie ;

Pelloux, Jerome ;

Lefebvre, Jean-Francois ;

Louvet, Romain ;

Rusterucci, Christine ;

Moritz, Thomas ;

Guerineau, Francois ;

Bellini, Catherine ;

Van Wuytswinkel, Olivier .

PLANT BIOTECHNOLOGY JOURNAL, 2008, 6 (06) :609-618

[7] qBase relative quantification framework and software for management and automated analysis of real-time quantitative PCR data [J].

Hellemans, Jan ;

Mortier, Geert ;

De Paepe, Anne ;

Speleman, Frank ;

Vandesompele, Jo .

GENOME BIOLOGY, 2007, 8 (02)

[8] Real-time RT-PCR normalisation; strategies and considerations [J].

Huggett, J ;

Dheda, K ;

Bustin, S ;

Zumla, A .

GENES AND IMMUNITY, 2005, 6 (04) :279-284

[9] Towards standardization of RNA quality assessment using user-independent classifiers of microcapillary electrophoresis traces [J].

Imbeaud, S ;

Graudens, E ;

Boulanger, V ;

Barlet, X ;

Zaborski, P ;

Eveno, E ;

Mueller, O ;

Schroeder, A ;

Auffray, C .

NUCLEIC ACIDS RESEARCH, 2005, 33 (06) :1-12

[10]

LEFEVER S, 2009, NUCL ACIDS RES 0217