Immobilization of oligonucleotides onto a glass support via disulfide bonds: A method for preparation of DNA microarrays.
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.
Rogers
Y H
YH
Alpha Center, Johns Hopkins Bayview Research Campus, 5210 Eastern Avenue, Baltimore, Maryland, 21224, USA.
Jiang-Baucom
P
P
Huang
Z J
ZJ
Bogdanov
V
V
Anderson
S
S
Boyce-Jacino
M T
MT
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
D015342:DNA Probes; D004220:Disulfides; D015345:Oligonucleotide Probes; D013438:Sulfhydryl Compounds