Nucleic Acids Research Advance Access originally published online on December 18, 2006
Nucleic Acids Research 2007 35(3):e15; doi:10.1093/nar/gkl1030
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Nucleic Acids Research, 2007, Vol. 35, No. 3 e15
© 2006 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.
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High resolution array-CGH analysis of single cells
The Wellcome Trust Sanger Institute, Wellcome Trust Genome Campus Hinxton, Cambridge CB10 1SA, UK 1 Institute of Medical Biology and Human Genetics, Medical University of Graz Harrachgasse 21/8, 8010 Graz, Austria 2 Institute of Human Genetics, Technical University Munich Trogerstrasse 32, 81675 Munich, Germany 3 Institute of Molecular Radiobiology, Molecular Cytogenetics, GSF-National Research Centre Ingolstaedter Landstrasse 1, 85764 Neuherberg, Germany
*To whom correspondence should be addressed. Tel: +44 1223 494842; Fax: +44 1223 494919; Email: heike{at}sanger.ac.uk
Received August 4, 2006. Revised September 28, 2006. Accepted November 15, 2006.
Heterogeneity in the genome copy number of tissues is of particular importance in solid tumor biology. Furthermore, many clinical applications such as pre-implantation and non-invasive prenatal diagnosis would benefit from the ability to characterize individual single cells. As the amount of DNA from single cells is so small, several PCR protocols have been developed in an attempt to achieve unbiased amplification. Many of these approaches are suitable for subsequent cytogenetic analyses using conventional methodologies such as comparative genomic hybridization (CGH) to metaphase spreads. However, attempts to harness array-CGH for single-cell analysis to provide improved resolution have been disappointing. Here we describe a strategy that combines single-cell amplification using GenomePlex library technology (GenomePlex® Single Cell Whole Genome Amplification Kit, Sigma-Aldrich, UK) and detailed analysis of genomic copy number changes by high-resolution array-CGH. We show that single copy changes as small as 8.3 Mb in single cells are detected reliably with single cells derived from various tumor cell lines as well as patients presenting with trisomy 21 and Prader–Willi syndrome. Our results demonstrate the potential of this technology for studies of tumor biology and for clinical diagnostics.
The authors wish to be known that, in their opinion, the first four authors should be regarded as joint First Authors
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