PMID: 9171075Jun 15, 1997Paper

Fractionation, phosphorylation and ligation on oligonucleotide microchips to enhance sequencing by hybridization

Nucleic Acids Research
S DubileyA D Mirzabekov

Abstract

Oligonucleotide microchips are manufactured by immobilizing presynthesized oligonucleotides within 0.1 x 0.1 x 0.02 mm or 1 x 1 x 0.02 mm polyacrylamide gel pads arranged on the surface of a microscope slide. The gel pads are separated from each other by hydrophobic glass spacers and serve as a kind of 'microtest tube' of 200 pl or 20 nl volume, respectively. Fractionation of single-stranded DNAs is carried out by their hybridization with chip pads containing immobilized 10mers. DNA extracted separately from each pad is transferred onto a sequencing chip and analyzed thereon. The chip, containing a set of 10mers, was enzymatically phosphorylated, then hybridized with DNA and ligated in a site-directed manner with a contiguously stacked 5mer. Several cycles of successive hybridization-ligation of the chip-bound 10mers with different contiguously stacked 5mers and hybridized with DNA were carried out to sequence DNA containing tetranucleotide repeats. Combined use of these techniques show significant promise for sequence comparison of homologous regions in different genomes and for sequence analysis of comparatively long DNA fragments or DNA containing internal repeats.

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Citations

Jul 14, 2005·Analytical and Bioanalytical Chemistry·Kenichi YamashitaHideaki Maeda
Apr 16, 1998·Current Opinion in Biotechnology·J H GraberC R Cantor
Feb 6, 2002·Current Opinion in Chemical Biology·Andrei Mirzabekov, Alexander Kolchinsky
Aug 6, 1998·Current Opinion in Chemical Biology·S C Case-GreenE M Southern
Jan 1, 2000·Journal of Biomolecular Structure & Dynamics·E KhomyakovaE Taillandier
Apr 29, 1998·Nucleic Acids Research·A V FotinA D Mirzabekov
Feb 7, 2001·Applied and Environmental Microbiology·S G BavykinA D Mirzabekov
Jul 20, 2010·Annual Review of Analytical Chemistry·Tim ConzeUlf Landegren
May 3, 2003·Analytical Sciences : the International Journal of the Japan Society for Analytical Chemistry·Sung Bae KimYoshio Umezawa
Feb 8, 2000·Analytical Biochemistry·P ArenkovA Mirzabekov
Apr 5, 2002·Human Mutation·Alexander Kolchinsky, Andrei Mirzabekov
Mar 1, 2008·Analytical Biochemistry·Zhiqiang PanZuhong Lu
Aug 16, 2005·Prostaglandins & Other Lipid Mediators·Li Feng
Jan 13, 2004·Analytical Biochemistry·A Yu RubinaA D Mirzabekov
Oct 3, 2000·Human Mutation·J G HaciaF S Collins
Aug 6, 2008·Biomacromolecules·Lu ChenJeffrey T Koberstein
Apr 2, 2002·Analytical Chemistry·Kimberly G OlsenMichael J Tarlov

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