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

Ericsson, Olle, Jarvius, Jonas, Schallmeiner, Edith, Howell, Mathias, Nong, Rachel Yuan, Reuter, Hendrik, Hahn, Meinhard, Stenberg, Johan, Nilsson, Mats, Landegren, Ulf
Nucleic acids research 2008 v.36 no.8 pp. e45
risk, proteins, messenger RNA, chemical elements, complementary DNA, monitoring, DNA microarrays, genes, nucleotide sequences
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.