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Quantitative real time PCR normalization and optimization

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.

Dr Joshua Yuan

  1. Content type: Research article

    Biomedical researchers have long looked for ways to diagnose and treat cancer patients at the early stages through biomarkers. Although conventional techniques are routinely applied in the detection of biomark...

    Authors: Keivan Majidzadeh-A, Rezvan Esmaeili and Nasrin Abdoli

    Citation: BMC Research Notes 2011 4:215

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  2. Content type: Short Report

    The use of RNAi to analyse gene function in vitro is now widely applied in biological research. However, several difficulties are associated with its use in vivo, mainly relating to inefficient delivery and non-s...

    Authors: Mike Herbert, Natacha Coppieters, Annette Lasham, Helen Cao and Glen Reid

    Citation: BMC Research Notes 2011 4:148

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  3. Content type: Technical Note

    The dog is frequently used as a model for hematologic human diseases. In this study the suitability of nine potential reference genes for quantitative RT-PCR studies in canine whole blood was investigated.

    Authors: Christine J Piek, Bas Brinkhof, Jan Rothuizen, Aldo Dekker and Louis C Penning

    Citation: BMC Research Notes 2011 4:36

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  4. Content type: Technical Note

    The efficiency of gene therapy experiments is frequently evaluated by measuring the impact of the treatment on the expression of genes of interest by quantitative real time PCR (qRT-PCR) and by normalizing the...

    Authors: María Moreno-Igoa, Raquel Manzano, Sara Oliván, Ana C Calvo, Janne M Toivonen and Rosario Osta

    Citation: BMC Research Notes 2010 3:275

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  5. Content type: Short Report

    Due to the limited number of species specific antibodies against fish proteins, differential gene expression analyses are vital for the study of host immune responses. Quantitative real-time reverse transcript...

    Authors: Andrea A Peña, Niels C Bols and Sergio H Marshall

    Citation: BMC Research Notes 2010 3:101

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  6. Content type: Short Report

    Rust caused by Puccinia psidii Winter has been limiting for the establishment of new Eucalyptus plantations, as well as for resprouting of susceptible genetic materials. Identifying host genes involved in defense...

    Authors: Leonardo P Boava, Marcelo L Laia, Tiago R Jacob, Karina M Dabbas, Janaína F Gonçalves, Jesus A Ferro, Maria IT Ferro and Edson L Furtado

    Citation: BMC Research Notes 2010 3:43

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  7. Content type: Project Note

    Reference genes are used as internal standards to normalize mRNA abundance in quantitative real-time PCR and thereby allow a direct comparison between samples. So far most of these expression studies used huma...

    Authors: Jan Axtner and Simone Sommer

    Citation: BMC Research Notes 2009 2:264

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  8. Content type: Short Report

    Application of reverse transcription quantitative real-time polymerase chain reaction is very well suited to reveal differences in gene expression between in vivo and in vitro produced embryos. Ultimately, this m...

    Authors: Katrien Smits, Karen Goossens, Ann Van Soom, Jan Govaere, Maarten Hoogewijs, Emilie Vanhaesebrouck, Cesare Galli, Silvia Colleoni, Jo Vandesompele and Luc Peelman

    Citation: BMC Research Notes 2009 2:246

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  9. Content type: Short Report

    The quantitative polymerase chain reaction (qPCR) is a widely utilized method for gene-expression analysis. However, insufficient material often compromises large-scale gene-expression studies. The aim of this...

    Authors: Joëlle Vermeulen, Stefaan Derveaux, Steve Lefever, Els De Smet, Katleen De Preter, Nurten Yigit, Anne De Paepe, Filip Pattyn, Frank Speleman and Jo Vandesompele

    Citation: BMC Research Notes 2009 2:235

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  10. Content type: Technical Note

    The development of reverse transcription – quantitative real-time PCR (RT-qPCR) platforms that can simultaneously measure the expression of multiple genes is dependent on robust assays that function under iden...

    Authors: Thomas Mikeska and Alexander Dobrovic

    Citation: BMC Research Notes 2009 2:112

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  11. Content type: Short Report

    We report the expression pattern of 5S rDNA in the eggs of water frogs Rana lessonae, Rana ridibunda and Rana esculenta using the quantitative real-time PCR. This kind of research had never been performed before.

    Authors: Elżbieta Czarniewska and Robert Plewa

    Citation: BMC Research Notes 2009 2:10

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  12. Content type: Short Report

    PCR inhibition by nucleic acid extracts is a well known yet poorly described phenomenon. Inhibition assessment generally depends on the assumption that inhibitors affect all PCR reactions to the same extent; i...

    Authors: Jim F Huggett, Tanya Novak, Jeremy A Garson, Clare Green, Stephen D Morris-Jones, Robert F Miller and Alimuddin Zumla

    Citation: BMC Research Notes 2008 1:70

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  13. Content type: Short Report

    Extensive sequencing efforts have been taking place for the Atlantic cod (Gadus morhua) in recent years, the number of ESTs in the Genbank has reached more than 140.000. Despite its importance in North Atlantic f...

    Authors: Pål A Olsvik, Liv Søfteland and Kai K Lie

    Citation: BMC Research Notes 2008 1:47

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    The Erratum to this article has been published in BMC Research Notes 2011 4:456

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