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Journal of Experimental & Clinical Cancer Research
Published in: Journal of Experimental & Clinical Cancer Research 1/2010

Open Access 01-12-2010 | Research

Evaluation of suitable reference genes for normalization of real-time reverse transcription PCR analysis in colon cancer

Authors: Lise Aa Sørby, Solveig N Andersen, Ida RK Bukholm, Morten B Jacobsen

Published in: Journal of Experimental & Clinical Cancer Research | Issue 1/2010

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Abstract

Background

Real-time reverse transcription PCR (qRT-PCR) is frequently used for gene expression quantification due to its methodological reproducibility and sensitivity. The gene expression is quantified by normalization to one or more reference genes which are presumed stably expressed throughout a given experiment. The aim of this study was to validate a standardized experimental setup to identifying reference genes for normalization of qRT-PCR in the metastatic and non-metastatic colon cancer.

Methods

In this study, expression of 16 commonly used reference genes was quantified in tumour tissue and individual-matched normal mucosa in 18 non-metastatic colon cancer patients and 20 colon cancer patients with distant metastases using TaqMan Low Density Array (TLDA). The expression stability was determined and compared by means of geNorm and NormFinder.

Results

Two pairs of genes, HPRT1/PPIA and IPO8/PPIA, were identified to be suitable to normalize gene expression data in metastatic and non-metastatic colon cancer patients, according to geNorm and NormFinder respectively.

Conclusion

We propose a standardized approach of finding the most suitable reference gene(s) in every qRT-PCR experiment using TLDA.
Appendix
Available only for authorised users
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Metadata
Title
Evaluation of suitable reference genes for normalization of real-time reverse transcription PCR analysis in colon cancer
Authors
Lise Aa Sørby
Solveig N Andersen
Ida RK Bukholm
Morten B Jacobsen
Publication date
01-12-2010
Publisher
BioMed Central
Published in
Journal of Experimental & Clinical Cancer Research / Issue 1/2010
Electronic ISSN: 1756-9966
DOI
https://doi.org/10.1186/1756-9966-29-144

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