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Nucleic Acids Research Advance Access published online on December 10, 2007
Nucleic Acids Research, doi:10.1093/nar/gkm1076
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© 2007 The Author(s)
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.

Methods Online

Polony analysis of gene expression in ES cells and blastocysts

C. Rieger1, R. Poppino1, R. Sheridan2, K. Moley2, R. Mitra3 and D. Gottlieb1,*

1Department of Anatomy and Neurobiology, 2Department of Obstetrics and Gynecology and 3Department of Genetics, Washington University School of Medicine, 660 S. Euclid Avenue, St Louis, MO 63110, USA

*To whom correspondence should be addressed. Tel: +314 362 2758; Fax: +314 362 3446; Email: gottlied{at}pcg.wustl.edu

Received June 28, 2007. Revised October 24, 2007. Accepted November 15, 2007.

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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