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Nucleic Acids Research, 2001, Vol. 29, No. 4 955-959
© 2001 Oxford University Press

Immobilization of oligodeoxyribonucleotides with multiple anchors to microchips

Xiaodong Zhao, Satyam Nampalli, Anthony J. Serino and Shiv Kumar*

Amersham Pharmacia Biotech Inc., 800 Centennial Avenue, Piscataway, NJ 08855, USA

Facile modification of oligodeoxyribonucleotides is required for efficient immobilization to a pre-activated glass surface. This report presents an oligodeoxyribonucleotide which contains a hairpin stem–loop structure with multiple phosphorothioate moieties in the loop. These moieties are used to anchor the oligo to glass slides that are pre-activated with bromoacetamidopropylsilane. The efficiency of the attachment reaction was improved by increasing the number of phosphorothioates in the loop, as shown in the remarkable enhancement of template hybridization and single base extension through catalysis by DNA polymerase. The loop and stem presumably serve as lateral spacers between neighboring oligodeoxyribonucleotides and as a linker arm between the glass surface and the single-stranded sequence of interest. The oligodeoxyribonucleotides of this hairpin stem–loop architecture with multiple phosphorothioate moieties have broad application in DNA chip-based gene analysis.

* To whom correspondence should be addressed. Tel: +1 732 980 2878; Fax: +1 732 457 8353; Email: shiv.kumar@am.apbiotech.com Present address: Xiaodong Zhao, Orchid BioSciences Inc., 303 College Road, Princeton, NJ 08540, USA


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