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Nucleic Acids Research Advance Access published online on March 16, 2008
Nucleic Acids Research, doi:10.1093/nar/gkn106
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© 2008 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

A dual-tag microarray platform for high-performance nucleic acid and protein analyses

Olle Ericsson1, Jonas Jarvius1, Edith Schallmeiner1, Mathias Howell1, Rachel Yuan Nong1, Hendrik Reuter2, Meinhard Hahn2, Johan Stenberg1, Mats Nilsson1 and Ulf Landegren*

1Department of Genetics and Pathology, The Rudbeck Laboratory, Uppsala University, SE-75185 Uppsala, Sweden and 2Division of Molecular Genetics, B060, Deutsches Krebsforschungszentrum, Im Neuenheimer Feld 280, D-69120 Heidelberg, Germany

*To whom correspondence should be addressed. Tel: +46 18 471 4910; Fax: +46 18 471 4808; Email: ulf.landegren{at}genpat.uu.se

Received December 5, 2007. Revised February 24, 2008. Accepted February 25, 2008.

DNA microarrays serve to monitor a wide range of molecular events, but emerging applications like measurements of weakly expressed genes or of proteins and their interaction patterns will require enhanced performance to improve specificity of detection and dynamic range. To further extend the utility of DNA microarray-based approaches we present a high-performance tag microarray procedure that enables probe-based analysis of as little as 100 target cDNA molecules, and with a linear dynamic range close to 105. Furthermore, the protocol radically decreases the risk of cross-hybridization on microarrays compared to current approaches, and it also allows for quantification by single-molecule analysis and real-time on-chip monitoring of rolling-circle amplification. We provide proof of concept for microarray-based measurement of both mRNA molecules and of proteins, converted to tag DNA sequences by padlock and proximity probe ligation, respectively.

The authors wish it to be known that, in their opinion, the first two authors should be regarded as joint First Authors.


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