A dual-tag microarray platform for high-performance nucleic acid and protein analyses.
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 10(5). 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.
Ericsson
Olle
O
Department of Genetics and Pathology, The Rudbeck Laboratory, Uppsala University, SE-75185 Uppsala, Sweden.
Jarvius
Jonas
J
Schallmeiner
Edith
E
Howell
Mathias
M
Nong
Rachel Yuan
RY
Reuter
Hendrik
H
Hahn
Meinhard
M
Stenberg
Johan
J
Nilsson
Mats
M
Landegren
Ulf
U
D000199:Actins / Q000235:genetics / Q000378:metabolism; D002460:Cell Line; D020869:Gene Expression Profiling / Q000379:methods*; D015335:Molecular Probes; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D010982:Platelet-Derived Growth Factor / Q000235:genetics / Q000378:metabolism; D016133:Polymerase Chain Reaction; D011506:Proteins / Q000032:analysis*; D012333:RNA, Messenger / Q000032:analysis*; D042461:Vascular Endothelial Growth Factor A / Q000032:analysis
D000199:Actins; D015335:Molecular Probes; D010982:Platelet-Derived Growth Factor; D011506:Proteins; D012333:RNA, Messenger; D042461:Vascular Endothelial Growth Factor A