Abstract
Allele-specific hybridization to a DNA microarray can be a useful method for genotyping patient DNA. In this article, we demonstrate that 13- to 17-base oligonucleotides tagged with a poly(T)10-poly(C)10 tail (TC tag), but otherwise unmodified, can be crosslinked by UV light irradiation to an agarose film grafted onto unmodified glass. Microarrays of TC-tagged probes immobilized on the agarose film can be used to diagnose mutations in the human beta-globin gene, which encodes the beta-chains in hemoglobin. Although the probes differed widely regarding melting point temperature ( approximately 20 degrees C), a single stringency wash still gave sufficiently high discrimination signals between perfect match and mismatch probes to allow robust mutation detection. In all, 270 genotypings were performed on patient materials, and no genotype was incorrectly classified. Quality control experiments conducted using a target DNA specific for the TC tag of the immobilized probes showed that the spotting and hybridization procedure had a variance of 20%, indicating that signal differences as low as twofold could be detected between perfect match and mismatch. Together, our results show that the use of microarrays of TC-tagged probes that ha...Continue Reading
References
Aug 1, 1989·Proceedings of the National Academy of Sciences of the United States of America·R K SaikiH A Erlich
Oct 25, 1996·Science·M CheeS P Fodor
Oct 1, 1996·Proceedings of the National Academy of Sciences of the United States of America·M SchenaR W Davis
Jun 6, 1998·Science·D G WangE S Lander
May 30, 1998·Analytical Biochemistry·D ProudnikovA Mirzabekov
Dec 24, 1998·Nucleic Acids Research·F N RehmanT C Boles
Jan 15, 1999·Analytical Biochemistry·Y H RogersM T Boyce-Jacino
Apr 13, 1999·Nucleic Acids Research·M Beier, J D Hoheisel
Jul 25, 2000·Nucleic Acids Research·A KumarZ Liang
Sep 13, 2000·Nucleic Acids Research·T StrotherL M Smith
May 18, 2001·Analytical Biochemistry·Y BelosludtsevM Hogan
Dec 26, 2001·Bioinformatics·N Le Novère
Jan 15, 2002·Nucleic Acids Research·Rüdiger BentersDieter Wöhrle
Jun 13, 2002·Nucleic Acids Research·Hong WangZuhong Lu
Feb 13, 2003·Nucleic Acids Research·Martin J HessnerSoumitra Ghosh
May 29, 2003·Nucleic Acids Research·Martin J HessnerSoumitra Ghosh
Jun 18, 2003·The Biochemical Journal·Sung-Kay ChiuChi-Meng Tzeng
Aug 9, 2003·Nucleic Acids Research·Véronique Le BerreJean François
Jan 14, 2004·Nucleic Acids Research·F FixeC B V Christensen
May 26, 2004·Lab on a Chip·F FixeC B V Christensen
Jun 17, 2005·Analytical Biochemistry·C A KochR S Utkhede
Oct 26, 2005·Biomolecular Engineering·Martin Dufva
Citations
Jun 6, 2009·Analytical and Bioanalytical Chemistry·Martin DufvaLena Poulsen
Sep 4, 2009·Analytical and Bioanalytical Chemistry·Thomas SchülerJürgen Popp
Oct 27, 2011·Analytical and Bioanalytical Chemistry·Yi SunAnders Wolff
Mar 26, 2010·Biomedical Microdevices·David SabourinMartin Dufva
Dec 8, 2007·Nucleic Acids Research·Jesper PetersenMartin Dufva
Sep 23, 2008·Nucleic Acids Research·Lena PoulsenMartin Dufva
Mar 12, 2009·PloS One·Jesper PetersenMartin Dufva
Mar 30, 2011·Biochemistry and Molecular Biology Education : a Bimonthly Publication of the International Union of Biochemistry and Molecular Biology·Thomas E Crowley
Sep 26, 2006·Analytical Biochemistry·Jesper PetersenMartin Dufva
Feb 22, 2017·Chemistry, an Asian Journal·Gyeongyeop HanJungkyu K Lee
Sep 10, 2008·BioTechniques·Haukur GudnasonAnders Wolff
Jul 16, 2014·International Journal of Oncology·Yuliang LiuAnders Wolff
Oct 24, 2007·Bioconjugate Chemistry·Naoki KimuraKoji Matsuoka