UNIT 12.6 Preparation of α-Oxo Semicarbazone Oligonucleotide Microarrays
Published Online: 1 JAN 2005
Copyright © 2004 by John Wiley and Sons, Inc.
Lab Protocol Title
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.
- Published Online: 1 JAN 2005
- Published Print: DEC 2004
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.
- α-oxo semicarbazone;
- α-oxo aldehyde;