Covalent attachment of DNA to glass supports using a new silane coupling agent and chemiluminescent detection

Abstract

A new kind of silane coupling agent, N-(β-aminoethyl)-γ-aminopropyl triethoxysilane, was used for DNA direct attachment on the surfaces of glass supports, then the immobilized DNA was hybridized with horseradish peroxidase (HRP)-labeled probe, and detected by using enhanced chemiluminescent method. In comparison with 7-aminopropyl triethoxysilane, the detection limits (S/N) of DNA were 10 pg and 75 pg respectively. Several experimental conditions of DNA attaching to glass supports were investigated, and the system of hybridization of nucleic acid on the surfaces of glass supports was developed.

Keywords

new silane coupling agent / DNA attachment / glass support / nucleic acid hybridization / enhanced chemiluminescent detection

References

[1] NikiforovT T, RendleR B, GoeletP, et al.. Genetic bit analysis: a solid phase method for typing single nucleotide polymorphisms. Nuc Acids Res, 1994, 22: 4167-4167

[2] ZammatteoN, GirardeauxC, DelforgeD, et al.. Amination of polystyrene microwells: application to the covalent grafting of DNA probes for hybridization assays. Anal Biochem, 1996, 236: 85-85

[3] RasmussenS R, LarsenM R, RasmussenS E.. Covalent immobilization of DNA onto polystyrene microwells: the molecules are only bound at the 5’ end. Anal Biochem, 1991, 198: 138-138

[4] MatsonR S, RampaiJ, PentoneyS L, et al.. Biopolymer synthesis on polypropylene supports: oligonucleotide array. Anal Biochem, 1995, 224: 110-110

[5] KhrapkoK R, LysovY P, IvanovI B, et al.. A method for DNA sequencing by hybridization with oligonucleotide matrix. J DNA Seq Map, 1991, 1: 375-375

[6] EggersM, HoganM, ReichR K, et al.. A microchip for quantitative detection of molecules utilizing luminescent and radioisotope reporter groups. Biotechniques, 1994, 17: 516-516

[7] HirschfeldT B. Device and method for nucleic acid analysis by hybridization assay with specifically binding fluorochrome. U. S. Patent 5, 1993, 242: 797-797

[8] JoosB, RusterH, ConeR, et al.. Covalent attachment of hybridizable oligonucleotides to glass supports. Anal Biochem, 1997, 247: 96-96

[9] 1997, 26(5):360