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Disease Markers
Volume 32, Issue 3, Pages 203-210
http://dx.doi.org/10.3233/DMA-2011-0875

Evaluation of Reference Genes and Normalization Strategy for Quantitative Real-Time PCR in Human Pancreatic Carcinoma

Beatrice Mohelnikova-Duchonova,1,2 Martin Oliverius,3 Eva Honsova,4 and Pavel Soucek1

1Department of Toxicogenomics, National Institute of Public Health, Prague, Czech Republic
2First Faculty of Medicine, Charles University in Prague, Czech Republic
3Department of Transplantation Surgery, Institute of Clinical and Experimental Medicine, Prague, Czech Republic
4Department of Clinical and Transplantation Pathology, Institute of Clinical and Experimental Medicine, Prague, Czech Republic

Received 23 February 2012; Accepted 23 February 2012

Copyright © 2012 Hindawi Publishing Corporation. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

Histologically verified pairs (n = 10) of pancreatic tumors and non-neoplastic tissues were used for quantitative real-time PCR and the stability of 24 reference genes was analyzed with geNorm and NormFinder software. Raw Cq values correlated with the degree of RNA degradation. This correlation was abolished by normalization to Cq of 18S endogenous control gene. Both geNorm and NormFinder programs suggested EIF2B1, ELF1, MRPL19, and POP4 as the same most stable genes. We have thus identified suitable reference genes for future expression studies in pancreatic carcinoma. Normalization method reducing the effects of RNA degradation on the quality of results was also developed.