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. 2012 Feb 29;32(3):203–210. doi: 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, Pavel Soucek 1,*
PMCID: PMC3826910  PMID: 22377737

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.

Keywords: Pancreas, carcinoma, transcript, quantification, reference gene, normalization

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