title: Immobilization of oligonucleotides onto a glass support via disulfide bonds: A method for preparation of DNA microarrays.
abstract: The covalent attachment of disulfide-modified oligonucleotides to a mercaptosilane-modified glass surface is described. This method provides an efficient and specific covalent attachment chemistry for immobilization of DNA probes onto a solid support. Glass slides were derivatized with 3-mercaptopropyl silane for attachment of 5-prime disulfide-modified oligonucleotides via disulfide bonds. An attachment density of approximately 3 x 10(5) oligonucleotides/microm2 was observed. Oligonucleotides attached by this method provided a highly efficient substrate for nucleic acid hybridization and primer extension assays. In addition, we have demonstrated patterning of multiple DNA probes on a glass surface utilizing this attachment chemistry, which allows for array densities of at least 20,000 spots/cm2.
authors.affiliation: Alpha Center, Johns Hopkins Bayview Research Campus, 5210 Eastern Avenue, Baltimore, Maryland, 21224, USA.
authors.forename: Y H
authors.initials: YH
authors.lastname: Rogers
authors.forename: P
authors.initials: P
authors.lastname: Jiang-Baucom
authors.forename: Z J
authors.initials: ZJ
authors.lastname: Huang
authors.forename: V
authors.initials: V
authors.lastname: Bogdanov
authors.forename: S
authors.initials: S
authors.lastname: Anderson
authors.forename: M T
authors.initials: MT
authors.lastname: Boyce-Jacino
mesh_terms: D015342:DNA Probes; D004220:Disulfides / Q000737:chemistry; D005898:Glass / Q000737:chemistry*; D006863:Hydrogen-Ion Concentration; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D015345:Oligonucleotide Probes / Q000737:chemistry*; D012680:Sensitivity and Specificity; D013438:Sulfhydryl Compounds / Q000737:chemistry; D013997:Time Factors
chemical_list: D015342:DNA Probes; D004220:Disulfides; D015345:Oligonucleotide Probes; D013438:Sulfhydryl Compounds