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Immobilization of oligonucleotides onto a glass support via disulfide bonds: A method for preparation of DNA
Y H Rogers1, P Jiang-Baucom, Z J Huang
1Alpha Center, Johns Hopkins Bayview Research Campus, 5210 Eastern Avenue, Baltimore, Maryland, 21224, USA.
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.