Unit

UNIT 12.6 Preparation of α-Oxo Semicarbazone Oligonucleotide Microarrays

  1. Oleg Melnyk1,
  2. Christophe Olivier1,
  3. Nathalie Ollivier1,
  4. Yves Lemoine2,
  5. David Hot2,
  6. Ludovic Huot2,
  7. Catherine Gouyette3

Published Online: 1 JAN 2005

DOI: 10.1002/0471142700.nc1206s19

Current Protocols in Nucleic Acid Chemistry

Current Protocols in Nucleic Acid Chemistry

How to Cite

Melnyk, O., Olivier, C., Ollivier, N., Lemoine, Y., Hot, D., Huot, L. and Gouyette, C. 2005. Preparation of α-Oxo Semicarbazone Oligonucleotide Microarrays. Current Protocols in Nucleic Acid Chemistry. 19:12.6:12.6.1–12.6.19.

Author Information

  1. 1

    Université de Lille 2, Lille, France

  2. 2

    Institut Pasteur de Lille, Lille, France

  3. 3

    Institut Pasteur, Paris, France

Publication History

  1. Published Online: 1 JAN 2005
  2. Published Print: DEC 2004

Abstract

This unit describes the preparation of α-oxo aldehyde functionalized oligodeoxynucleotides, the preparation and characterization of semicarbazide glass slides, and the fabrication of α-oxo semicarbazone microarrays by site-specific ligation of α-oxo-aldehyde oligodeoxynucleotides to the semicarbazide glass slides. The α-oxo aldehyde group COCHO is extensively used in ligation chemistry for the preparation of large molecular constructs. It is stable toward air oxidation and mainly present in aqueous solution in the hydrated form COC(OH)2. It reacts efficiently with hydrazine derivatives, in particular, with the semicarbazide group. The reaction occurs spontaneously in water at pH 5.5. Site-specific immobilization of glyoxylyl oligodeoxynucleotides on semicarbazide glass slides allows the preparation of high-quality microarrays that can be used directly in hybridization experiments.

Keywords:

  • microarray;
  • oligonucleotide;
  • α-oxo semicarbazone;
  • α-oxo aldehyde;
  • semicarbazide