High surface density immobilization of oligonucleotide on silicon

The Analyst
B A CavicM Thompson

Abstract

Oligonucleotide (11-mer) molecules are immobilized on silicon in high surface population using either a permanent thioether bond or a chemo-selectively reversible disulfide bond to the surface thiol functionality. Substrate hydroxy groups are first silanized with an 11 carbon trichlorosilane containing a terminal, protected thiol moiety. Oligonucleotide modified with a tether possessing a terminal thiol group is further derivatized with a water-soluble, halobenzylic bifunctional reagent, which allows the complete conjugate to be attached to the surface through a permanent thioether bond. Alternatively, the oligonucleotide-tether complex can be combined with a pyridyldisulfide compound, which, in turn, facilitates the formation of a reversible disulfide bond with surface thiol. The amount of immobilized oligonucleotide was determined by radiochemical labeling with 32P. Additional verification of surface amounts was obtained from X-ray photoelectron spectroscopic analysis of substrates. The results of the immobilization protocols are compared with the oligonucleotide surface population achieved through the conventional silanizing agent, mercaptopropyltrimethoxysilane. Finally, a preliminary confirmation of duplex formation of a T...Continue Reading

Citations

Oct 31, 2006·Omics : a Journal of Integrative Biology·Soon Jin OhJoon Won Park
Sep 25, 2007·Biosensors & Bioelectronics·Sanjun NiuRavi F Saraf
Aug 6, 2008·Biomacromolecules·Lu ChenJeffrey T Koberstein
Sep 24, 2004·Langmuir : the ACS Journal of Surfaces and Colloids·Dae-Hwan JungHee-Tae Jung
Dec 13, 2006·Langmuir : the ACS Journal of Surfaces and Colloids·Jung-Hoon YangHiroshi Kawarada

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