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

• Published in: Nucleic acids research Vol. 36; no. 8; p. e45
• Main Authors: Ericsson, Olle, Jarvius, Jonas, Schallmeiner, Edith, Howell, Mathias, Nong, Rachel Yuan, Reuter, Hendrik, Hahn, Meinhard, Stenberg, Johan, Nilsson, Mats, Landegren, Ulf
• Format: Journal Article
• Language: English
• Published: England Oxford University Press 01.05.2008; Oxford Publishing Limited (England)
• Subjects: Actins - genetics; Actins - metabolism; Cell Line; Gene Expression Profiling - methods; MEDICIN; MEDICINE; Methods Online; Molecular Probes; Oligonucleotide Array Sequence Analysis - methods; Platelet-Derived Growth Factor - genetics; Platelet-Derived Growth Factor - metabolism; Polymerase Chain Reaction; Proteins - analysis; RNA, Messenger - analysis; Vascular Endothelial Growth Factor A - analysis
• ISSN: 0305-1048; 1362-4962; 1362-4962
• DOI: 10.1093/nar/gkn106

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.