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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. 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
    Content type: Research article Published on:

  2. 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
    Content type: Short Report Published on:

  3. 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
    Content type: Technical Note Published on:

  4. 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
    Content type: Technical Note Published on:

  5. 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
    Content type: Short Report Published on:

  6. 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
    Content type: Short Report Published on:

  7. 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
    Content type: Project Note Published on:

  8. 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
    Content type: Short Report Published on:

  9. 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
    Content type: Short Report Published on:

  10. 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
    Content type: Technical Note Published on:

  11. 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
    Content type: Short Report Published on:

  13. 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
    Content type: Short Report Published on:

  14. 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
    Content type: Short Report Published on:

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