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Selection of Candidate Housekeeping Genes for Normalization in Human Postmortem Brain Samples

The most frequently used technique to study the expression profile of genes involved in common neurological disorders is quantitative real-time RT-PCR, which allows the indirect detection of very low amounts of selected mRNAs in tissue samples. Expression analysis by RT-qPCR requires an appropriate...

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Detalles Bibliográficos
Autores principales: Penna, Ilaria, Vella, Serena, Gigoni, Arianna, Russo, Claudio, Cancedda, Ranieri, Pagano, Aldo
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Molecular Diversity Preservation International (MDPI) 2011
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3189726/
https://www.ncbi.nlm.nih.gov/pubmed/22016602
http://dx.doi.org/10.3390/ijms12095461
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author Penna, Ilaria
Vella, Serena
Gigoni, Arianna
Russo, Claudio
Cancedda, Ranieri
Pagano, Aldo
author_facet Penna, Ilaria
Vella, Serena
Gigoni, Arianna
Russo, Claudio
Cancedda, Ranieri
Pagano, Aldo
author_sort Penna, Ilaria
collection PubMed
description The most frequently used technique to study the expression profile of genes involved in common neurological disorders is quantitative real-time RT-PCR, which allows the indirect detection of very low amounts of selected mRNAs in tissue samples. Expression analysis by RT-qPCR requires an appropriate normalization to the expression level of genes characterized by a stable, constitutive transcription. However, the identification of a gene transcribed at a very stable level is difficult if not impossible, since significant fluctuations of the level of mRNA synthesis often accompanies changes of cell behavior. The aim of this study is to identify the most stable genes in postmortem human brain samples of patients affected by Alzheimer’s disease (AD) suitable as reference genes. The experiments analyzed 12 commonly used reference genes in brain samples from eight individuals with AD and seven controls. After a careful analysis of the results calculated by geNorm and NormFinder algorithms, we found that CYC1 and EIF4A2 are the best reference genes. We remark on the importance of the determination of the best reference genes for each sample to be analyzed and suggest a practical combination of reference genes to be used in the analysis of human postmortem samples.
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spelling pubmed-31897262011-10-20 Selection of Candidate Housekeeping Genes for Normalization in Human Postmortem Brain Samples Penna, Ilaria Vella, Serena Gigoni, Arianna Russo, Claudio Cancedda, Ranieri Pagano, Aldo Int J Mol Sci Communication The most frequently used technique to study the expression profile of genes involved in common neurological disorders is quantitative real-time RT-PCR, which allows the indirect detection of very low amounts of selected mRNAs in tissue samples. Expression analysis by RT-qPCR requires an appropriate normalization to the expression level of genes characterized by a stable, constitutive transcription. However, the identification of a gene transcribed at a very stable level is difficult if not impossible, since significant fluctuations of the level of mRNA synthesis often accompanies changes of cell behavior. The aim of this study is to identify the most stable genes in postmortem human brain samples of patients affected by Alzheimer’s disease (AD) suitable as reference genes. The experiments analyzed 12 commonly used reference genes in brain samples from eight individuals with AD and seven controls. After a careful analysis of the results calculated by geNorm and NormFinder algorithms, we found that CYC1 and EIF4A2 are the best reference genes. We remark on the importance of the determination of the best reference genes for each sample to be analyzed and suggest a practical combination of reference genes to be used in the analysis of human postmortem samples. Molecular Diversity Preservation International (MDPI) 2011-08-26 /pmc/articles/PMC3189726/ /pubmed/22016602 http://dx.doi.org/10.3390/ijms12095461 Text en © 2011 by the authors; licensee MDPI, Basel, Switzerland. http://creativecommons.org/licenses/by/3.0 This article is an open-access article distributed under the terms and conditions of the Creative Commons Attribution license (http://creativecommons.org/licenses/by/3.0/).
spellingShingle Communication
Penna, Ilaria
Vella, Serena
Gigoni, Arianna
Russo, Claudio
Cancedda, Ranieri
Pagano, Aldo
Selection of Candidate Housekeeping Genes for Normalization in Human Postmortem Brain Samples
title Selection of Candidate Housekeeping Genes for Normalization in Human Postmortem Brain Samples
title_full Selection of Candidate Housekeeping Genes for Normalization in Human Postmortem Brain Samples
title_fullStr Selection of Candidate Housekeeping Genes for Normalization in Human Postmortem Brain Samples
title_full_unstemmed Selection of Candidate Housekeeping Genes for Normalization in Human Postmortem Brain Samples
title_short Selection of Candidate Housekeeping Genes for Normalization in Human Postmortem Brain Samples
title_sort selection of candidate housekeeping genes for normalization in human postmortem brain samples
topic Communication
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3189726/
https://www.ncbi.nlm.nih.gov/pubmed/22016602
http://dx.doi.org/10.3390/ijms12095461
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