DNA Microarrays Based on Noncovalent Oligonucleotide Attachment and Hybridization in Two Dimensions

✍ Scribed by Yuri Belosludtsev; Bonnie Iverson; Sergey Lemeshko; Rick Eggers; Rick Wiese; Sandy Lee; Tom Powdrill; Mike Hogan

• Book ID: 102567274
• Publisher: Elsevier Science
• Year: 2001
• Tongue: English
• Weight: 118 KB
• Volume: 292
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1006/abio.2001.5088

✦ Synopsis

Short oligonucleotide probes have been linked to a solid support by simple electrostatic adsorption onto a positively charged surface film. Attachment was obtained by microfluidic application of unmodified oligonucleotides in distilled water onto amino-silanized glass. It has been demonstrated that an extremely stable monolayer of oligonucleotide is obtained by this method, at a density of about 10 11 molecules/mm 2 , which approaches the limit expected for a two-dimensional closest-packed array. Application of oligonucleotide by adsorption is followed by capping with acetic anhydride in the vapor phase, and then capping with succinic anhydride in solution to form a surface with weak negative charge. The capping method has been successfully employed for microarray fabrication and for the analysis of single nucleotide polymorphisms in the k-ras gene. The data reveal that, subsequent to capping, the adsorptive association of oligonucleotide to the surface yields a probe layer which is capable of single nucleotide base mismatch discrimination and high apparent binding affinity.