Oligonucleotide microarray preparation using enhanced poly-L-lysine glass slides

Abstract

Abstract: Objective To modify conventional poly-L-lysine coating for oligonucleotide microarray preparation so as to enhance the sensitivity of the microarray.Methods The proposed chemical approaches included silanizing the slides with 3-glycid-oxypropyltrimethoxysilane (GOPS) after cleanling, followed by slide coating with polymers (poly-L-lysine) that was covalently bound to the modified glass. Subsequent attachment of the oligonucleotide to the modified slide surface was achieved after 1,4-phenylene diisothiocyanate (PDITC) activation of the surface. Various experiments were carried out, such as the immobilization efficiency and hybridization assays to test the modified slides, which were then used tentatively in the preparation of microarrays for SARS coronavirus detection.Results The improved surface had high immobilization efficiency, good uniformity and satisfactory hybridization efficiency, better than those slides with conventional poly-L-lysinecoating. In addition, such modified slides rendered the microarrays more resistant to consecutive probing/stripping cycles.Conclusion The modified slide surface is satisfactory to immobilize unmodified oligonucleotide by covalent binding, which enhances not only the sensitivity of the prepared oligonucleotide microarray but also the binding of the oligonucleotide to the slide surface.

CLC Number:

WU Qing-hua¹, MA Wen-li¹, SHI Rong¹, GUO Qiu-ye¹, ZHANG Bao¹, LI Ling¹, ZHANG Hai-yan¹, ZHENG Wen-ling². Oligonucleotide microarray preparation using enhanced poly-L-lysine glass slides[J]. Journal of Southern Medical University, 2004, 24(11): 1236-1241.

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