Quantitative real time PCR normalization and optimization
Edited by:
Dr Joshua Yuan
Fluorescence-based quantitative real-time PCR (qRT-PCR) is a widely and commonly used technology to quantify DNA and RNA products. The main applications of qRT-PCR are diagnostic for rapid detection of nucleic acids characteristic of infectious diseases, cancer or genetic abnormalities and, when coupled with reverse transcription, it is mainly used to provide quantitative measurements of gene transcription. The validity and reproducibility of the results of qRT-PCR studies depend on numerous factors, including but not limited to, adequate reporting of experimental settings, choice of appropriate reference genes and statistical analysis of the data generated. This topical series is a collection of manuscripts relevant to researchers interested in normalization and validation of qRT-PCR experiments.
Collection published: 20 October 2008
Last updated: 25 June 2011
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TFRC and ACTB as the best reference genes to quantify Urokinase Plasminogen Activator in breast cancer
Keivan Majidzadeh-A, Rezvan Esmaeili, Nasrin Abdoli BMC Research Notes 2011, 4:215 (25 June 2011)
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The importance of RT-qPCR primer design for the detection of siRNA-mediated mRNA silencing
Mike Herbert, Natacha Coppieters, Annette Lasham, Helen Cao, Glen Reid BMC Research Notes 2011, 4:148 (25 May 2011)
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Leukocyte count affects expression of reference genes in canine whole blood samples
Christine J Piek, Bas Brinkhof, Jan Rothuizen, Aldo Dekker, Louis C Penning BMC Research Notes 2011, 4:36 (9 February 2011)
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Effects of gene therapy on muscle 18S rRNA expression in mouse model of ALS
María Moreno-Igoa, Raquel Manzano, Sara Oliván, Ana C Calvo, Janne M Toivonen, Rosario Osta BMC Research Notes 2010, 3:275 (2 November 2010)
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An evaluation of potential reference genes for stability of expression in two salmonid cell lines after infection with either Piscirickettsia salmonis or IPNV
Andrea A Peña, Niels C Bols, Sergio H Marshall BMC Research Notes 2010, 3:101 (14 April 2010)
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| PubMed | Cited on BioMed Central
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Selection of endogenous genes for gene expression studies in Eucalyptus under biotic (Puccinia psidii) and abiotic (acibenzolar-S-methyl) stresses using RT-qPCR
Leonardo P Boava, Marcelo L Laia, Tiago R Jacob, Karina M Dabbas, Janaína F Gonçalves, Jesus A Ferro, Maria IT Ferro, Edson L Furtado BMC Research Notes 2010, 3:43 (24 February 2010)
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Validation of internal reference genes for quantitative real-time PCR in a non-model organism, the yellow-necked mouse, Apodemus flavicollis
Jan Axtner, Simone Sommer BMC Research Notes 2009, 2:264 (23 December 2009)
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Selection of reference genes for quantitative real-time PCR in equine in vivo and fresh and frozen-thawed in vitro blastocysts
Katrien Smits, Karen Goossens, Ann Van Soom, Jan Govaere, Maarten Hoogewijs, Emilie Vanhaesebrouck, Cesare Galli, Silvia Colleoni, Jo Vandesompele, Luc Peelman BMC Research Notes 2009, 2:246 (11 December 2009)
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RNA pre-amplification enables large-scale RT-qPCR gene-expression studies on limiting sample amounts
Joëlle Vermeulen, Stefaan Derveaux, Steve Lefever, Els De Smet, Katleen De Preter, Nurten Yigit, Anne De Paepe, Filip Pattyn, Frank Speleman, Jo Vandesompele BMC Research Notes 2009, 2:235 (25 November 2009)
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Validation of a primer optimisation matrix to improve the performance of reverse transcription – quantitative real-time PCR assays
Thomas Mikeska, Alexander Dobrovic BMC Research Notes 2009, 2:112 (23 June 2009)
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Expression of 5S rDNA in the oocytes of water frogs
Elżbieta Czarniewska, Robert Plewa BMC Research Notes 2009, 2:10 (19 January 2009)
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DNA-free RNA isolation protocols for Arabidopsis thaliana, including seeds and siliques
Luis Oñate-Sánchez, Jesús Vicente-Carbajosa BMC Research Notes 2008, 1:93 (20 October 2008)
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Differential susceptibility of PCR reactions to inhibitors: an important and unrecognised phenomenon
Jim F Huggett, Tanya Novak, Jeremy A Garson, Clare Green, Stephen D Morris-Jones, Robert F Miller, Alimuddin Zumla BMC Research Notes 2008, 1:70 (28 August 2008)
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Selection of reference genes for qRT-PCR examination of wild populations of Atlantic cod Gadus morhua
Pål A Olsvik, Liv Søfteland, Kai K Lie BMC Research Notes 2008, 1:47 (16 July 2008)
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