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The correlation between expression profiles measured in single cells and in traditional bulk samples

Reverse transcription quantitative PCR (RT-qPCR) is already an established tool for mRNA detection and quantification. Since recently, this technique has been successfully employed for gene expression analyses, and also in individual cells (single cell RT-qPCR). Although the advantages of single cel...

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Autores principales: Dzamba, David, Valihrach, Lukas, Kubista, Mikael, Anderova, Miroslava
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5111061/
https://www.ncbi.nlm.nih.gov/pubmed/27848982
http://dx.doi.org/10.1038/srep37022
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author Dzamba, David
Valihrach, Lukas
Kubista, Mikael
Anderova, Miroslava
author_facet Dzamba, David
Valihrach, Lukas
Kubista, Mikael
Anderova, Miroslava
author_sort Dzamba, David
collection PubMed
description Reverse transcription quantitative PCR (RT-qPCR) is already an established tool for mRNA detection and quantification. Since recently, this technique has been successfully employed for gene expression analyses, and also in individual cells (single cell RT-qPCR). Although the advantages of single cell measurements have been proven several times, a study correlating the expression measured on single cells, and in bulk samples consisting of a large number of cells, has been missing. Here, we collected a large data set to explore the relation between gene expression measured in single cells and in bulk samples, reflected by qPCR Cq values. We measured the expression of 95 genes in 12 bulk samples, each containing thousands of astrocytes, and also in 693 individual astrocytes. Combining the data, we described the relation between Cq values measured in bulk samples with either the percentage of the single cells that express the given genes, or the average expression of the genes across the single cells. We show that data obtained with single cell RT-qPCR are fully consistent with measurements in bulk samples. Our results further provide a base for quality control in single cell expression profiling, and bring new insights into the biological process of cellular expression.
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spelling pubmed-51110612016-11-23 The correlation between expression profiles measured in single cells and in traditional bulk samples Dzamba, David Valihrach, Lukas Kubista, Mikael Anderova, Miroslava Sci Rep Article Reverse transcription quantitative PCR (RT-qPCR) is already an established tool for mRNA detection and quantification. Since recently, this technique has been successfully employed for gene expression analyses, and also in individual cells (single cell RT-qPCR). Although the advantages of single cell measurements have been proven several times, a study correlating the expression measured on single cells, and in bulk samples consisting of a large number of cells, has been missing. Here, we collected a large data set to explore the relation between gene expression measured in single cells and in bulk samples, reflected by qPCR Cq values. We measured the expression of 95 genes in 12 bulk samples, each containing thousands of astrocytes, and also in 693 individual astrocytes. Combining the data, we described the relation between Cq values measured in bulk samples with either the percentage of the single cells that express the given genes, or the average expression of the genes across the single cells. We show that data obtained with single cell RT-qPCR are fully consistent with measurements in bulk samples. Our results further provide a base for quality control in single cell expression profiling, and bring new insights into the biological process of cellular expression. Nature Publishing Group 2016-11-16 /pmc/articles/PMC5111061/ /pubmed/27848982 http://dx.doi.org/10.1038/srep37022 Text en Copyright © 2016, The Author(s) http://creativecommons.org/licenses/by/4.0/ This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/
spellingShingle Article
Dzamba, David
Valihrach, Lukas
Kubista, Mikael
Anderova, Miroslava
The correlation between expression profiles measured in single cells and in traditional bulk samples
title The correlation between expression profiles measured in single cells and in traditional bulk samples
title_full The correlation between expression profiles measured in single cells and in traditional bulk samples
title_fullStr The correlation between expression profiles measured in single cells and in traditional bulk samples
title_full_unstemmed The correlation between expression profiles measured in single cells and in traditional bulk samples
title_short The correlation between expression profiles measured in single cells and in traditional bulk samples
title_sort correlation between expression profiles measured in single cells and in traditional bulk samples
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5111061/
https://www.ncbi.nlm.nih.gov/pubmed/27848982
http://dx.doi.org/10.1038/srep37022
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