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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...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Molecular Diversity Preservation International (MDPI)
2011
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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. |
format | Online Article Text |
id | pubmed-3189726 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2011 |
publisher | Molecular Diversity Preservation International (MDPI) |
record_format | MEDLINE/PubMed |
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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