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Optimizing processing parameters for signal enhancement of oligonucleotide and protein arrays on ARChip Epoxy by C. Preininger; U. Sauer; J. Dayteg; R. Pichler is a Engineering article available to read on EtoBox.

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Signal enhancement of oligonucleotide and protein arrays on ARChip Epoxy was achieved by optimizing chip processing parameters. The parameters investigated were fabrication, blocking and guide dot concentration, probe concentration and modification, print buffer, humidity during arraying, slide agitation, spot volume and spotter compatibility. The optimum oligonucleotide concentration was 20 microM, while the optimum protein concentration was 0.05 mg/ml. Amino-modified oligonucleotides were best able to be bound to the resin's epoxy groups at pH 8, whereas thiol-modified oligonucleotides displayed an optimum coupling value of pH 7. So as to avoid background (BG) contamination of probes around bright guide dots, the concentration of fluorescent guide dots was set to 1 muM. The most suitable print buffers for oligonucleotide arrays using both piezo- and contact-printing systems proved to be 3 x SSC/1.5 M betaine and commercial ArrayLink. When 0.01% monochlortriazinyl-beta-cyclodextrin sodium salt (MCT) was added, the hybridization signal doubled in strength as compared to plain buffer. The optimum print buffer for proteins was 0.1 N phosphate buffer, pH 8/10% glycerine. The optimum h

Who reads Optimizing processing parameters for signal enhancement of oligonucleotide and protein arrays on ARChip Epoxy?

Author

C. Preininger; U. Sauer; J. Dayteg; R. Pichler

Publisher

Elsevier Science; Elsevier ; Elsevier BV (ISSN 1567-5394)

Published

2005

Language

EN

Field

Engineering (Physical Sciences)