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Polony analysis of gene expression in ES cells and blastocysts

Expression profiling of stem cells is challenging due to their small numbers and heterogeneity. The PCR colony (polony) approach has theoretical advantages as an assay for stem cells but has not been applied to small numbers of cells. An assay has been developed that is sensitive enough to detect mR...

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Autores principales: Rieger, C., Poppino, R., Sheridan, R., Moley, K., Mitra, R., Gottlieb, D.
Formato: Texto
Lenguaje:English
Publicado: Oxford University Press 2007
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2190707/
https://www.ncbi.nlm.nih.gov/pubmed/18073198
http://dx.doi.org/10.1093/nar/gkm1076
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author Rieger, C.
Poppino, R.
Sheridan, R.
Moley, K.
Mitra, R.
Gottlieb, D.
author_facet Rieger, C.
Poppino, R.
Sheridan, R.
Moley, K.
Mitra, R.
Gottlieb, D.
author_sort Rieger, C.
collection PubMed
description Expression profiling of stem cells is challenging due to their small numbers and heterogeneity. The PCR colony (polony) approach has theoretical advantages as an assay for stem cells but has not been applied to small numbers of cells. An assay has been developed that is sensitive enough to detect mRNAs from small numbers of ES cells and from fractions of a single mouse blastocyst. Genes assayed include Oct3, Rex1, Nanog, Cdx2 and GLUT-1. The assay is highly sensitive so that multiple mRNAs from a single blastocyst were easily detected in the same assay. In its present version, the assay is an attractive alternative to conventional RT–PCR for profiling small populations of stem cells. The assay is also amenable to improvements that will increase its sensitivity and ability to analyze many cDNAs simultaneously.
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spelling pubmed-21907072008-01-25 Polony analysis of gene expression in ES cells and blastocysts Rieger, C. Poppino, R. Sheridan, R. Moley, K. Mitra, R. Gottlieb, D. Nucleic Acids Res Methods Online Expression profiling of stem cells is challenging due to their small numbers and heterogeneity. The PCR colony (polony) approach has theoretical advantages as an assay for stem cells but has not been applied to small numbers of cells. An assay has been developed that is sensitive enough to detect mRNAs from small numbers of ES cells and from fractions of a single mouse blastocyst. Genes assayed include Oct3, Rex1, Nanog, Cdx2 and GLUT-1. The assay is highly sensitive so that multiple mRNAs from a single blastocyst were easily detected in the same assay. In its present version, the assay is an attractive alternative to conventional RT–PCR for profiling small populations of stem cells. The assay is also amenable to improvements that will increase its sensitivity and ability to analyze many cDNAs simultaneously. Oxford University Press 2007-12 2007-12-10 /pmc/articles/PMC2190707/ /pubmed/18073198 http://dx.doi.org/10.1093/nar/gkm1076 Text en © 2007 The Author(s) http://creativecommons.org/licenses/by-nc/2.0/uk/ This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Methods Online
Rieger, C.
Poppino, R.
Sheridan, R.
Moley, K.
Mitra, R.
Gottlieb, D.
Polony analysis of gene expression in ES cells and blastocysts
title Polony analysis of gene expression in ES cells and blastocysts
title_full Polony analysis of gene expression in ES cells and blastocysts
title_fullStr Polony analysis of gene expression in ES cells and blastocysts
title_full_unstemmed Polony analysis of gene expression in ES cells and blastocysts
title_short Polony analysis of gene expression in ES cells and blastocysts
title_sort polony analysis of gene expression in es cells and blastocysts
topic Methods Online
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2190707/
https://www.ncbi.nlm.nih.gov/pubmed/18073198
http://dx.doi.org/10.1093/nar/gkm1076
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