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

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 C(q) values correlated with the degree of RNA degradation. This correlation wa...

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Autores principales: Mohelnikova-Duchonova, Beatrice, Oliverius, Martin, Honsova, Eva, Soucek, Pavel
Formato: Online Artículo Texto
Lenguaje:English
Publicado: IOS Press 2012
Materias:
Other
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3826910/
https://www.ncbi.nlm.nih.gov/pubmed/22377737
http://dx.doi.org/10.3233/DMA-2011-0875
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author Mohelnikova-Duchonova, Beatrice
Oliverius, Martin
Honsova, Eva
Soucek, Pavel
author_facet Mohelnikova-Duchonova, Beatrice
Oliverius, Martin
Honsova, Eva
Soucek, Pavel
author_sort Mohelnikova-Duchonova, Beatrice
collection PubMed
description 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 C(q) values correlated with the degree of RNA degradation. This correlation was abolished by normalization to C(q) 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.
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spelling pubmed-38269102013-12-04 Evaluation of Reference Genes and Normalization Strategy for Quantitative Real-Time PCR in Human Pancreatic Carcinoma Mohelnikova-Duchonova, Beatrice Oliverius, Martin Honsova, Eva Soucek, Pavel Dis Markers Other 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 C(q) values correlated with the degree of RNA degradation. This correlation was abolished by normalization to C(q) 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. IOS Press 2012 2012-02-29 /pmc/articles/PMC3826910/ /pubmed/22377737 http://dx.doi.org/10.3233/DMA-2011-0875 Text en Copyright © 2012 Hindawi Publishing Corporation.
spellingShingle Other
Mohelnikova-Duchonova, Beatrice
Oliverius, Martin
Honsova, Eva
Soucek, Pavel
Evaluation of Reference Genes and Normalization Strategy for Quantitative Real-Time PCR in Human Pancreatic Carcinoma
title Evaluation of Reference Genes and Normalization Strategy for Quantitative Real-Time PCR in Human Pancreatic Carcinoma
title_full Evaluation of Reference Genes and Normalization Strategy for Quantitative Real-Time PCR in Human Pancreatic Carcinoma
title_fullStr Evaluation of Reference Genes and Normalization Strategy for Quantitative Real-Time PCR in Human Pancreatic Carcinoma
title_full_unstemmed Evaluation of Reference Genes and Normalization Strategy for Quantitative Real-Time PCR in Human Pancreatic Carcinoma
title_short Evaluation of Reference Genes and Normalization Strategy for Quantitative Real-Time PCR in Human Pancreatic Carcinoma
title_sort evaluation of reference genes and normalization strategy for quantitative real-time pcr in human pancreatic carcinoma
topic Other
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3826910/
https://www.ncbi.nlm.nih.gov/pubmed/22377737
http://dx.doi.org/10.3233/DMA-2011-0875
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