Gene Expression Analysis Using Oligonucleotide Arrays Produced by Maskless Photolithography

Author:

Nuwaysir Emile F.,Huang Wei,Albert Thomas J.,Singh Jaz,Nuwaysir Kate,Pitas Alan,Richmond Todd,Gorski Tom,Berg James P.,Ballin Jeff,McCormick Mark,Norton Jason,Pollock Tim,Sumwalt Terry,Butcher Lawrence,Porter DeAnn,Molla Michael,Hall Christine,Blattner Fred,Sussman Michael R.,Wallace Rodney L.,Cerrina Franco,Green Roland D.

Abstract

Microarrays containing 195,000 in situ synthesized oligonucleotide features have been created using a benchtop, maskless photolithographic instrument. This instrument, the Maskless Array Synthesizer (MAS), uses a digital light processor (DLP) developed by Texas Instruments. The DLP creates the patterns of UV light used in the light-directed synthesis of oligonucleotides. This digital mask eliminates the need for expensive and time-consuming chromium masks. In this report, we describe experiments in which we tested this maskless technology for DNA synthesis on glass surfaces. Parameters examined included deprotection rates, repetitive yields, and oligonucleotide length. Custom gene expression arrays were manufactured and hybridized toDrosophila melanogaster and mouse samples. Quantitative PCR was used to validate the gene expression data from the mouse arrays.[The sequence data from this study have been submitted to GEO under accession nos. GPL208, GSM2409, GSM2410, GSM2411, GSM2412, GSM2413, GSM2414, GSE81, GSE82.]

Publisher

Cold Spring Harbor Laboratory

Subject

Genetics (clinical),Genetics

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