Attachment of oligonucleotide probes to poly carbodiimide-coated glass for microarray applications
Open Access
- 1 April 2004
- journal article
- research article
- Published by Oxford University Press (OUP) in Nucleic Acids Research
- Vol. 32 (7) , e68
- https://doi.org/10.1093/nar/gnh057
Abstract
Oligonucleotide-based DNA microarrays are becoming increasingly useful tools for the analysis of gene expression and single nucleotide polymorphisms (SNPs). Here, we present a method that permits the manufacture of microarrays from non-modified oligonucleotides on a poly carbodiimide-coated glass surface by UV-irradiation. The use of UV-irradiation facilitates an increase in the level of signal intensity, but it does not affect signal discrimination by the oligonucleotides immobilized on the surface. The signal intensity obtained for an array fabricated using non-modified oligonucleotides with UV-irradiation is ∼7-fold greater than that without UV-irradiation. The detection of SNPs was tested to ascertain whether this technique could discriminate specific hybridization signals without causing significant UV-irradiation-induced damage to the immobilized oligonucleotides. We found that this immobilization method provides greater hybridization signals and a better match/mismatch ratio of SNPs than do the established aminosilane techniques. Application of this technology to manufacturing DNA microarrays for sequence analysis is discussed.Keywords
This publication has 22 references indexed in Scilit:
- Selection of oligonucleotide probes for protein coding sequencesBioinformatics, 2003
- G protein ??3 subunit 825T allele is associated with depression in young, healthy subjectsNeuroReport, 2003
- Assessing unmodified 70-mer oligonucleotide probe performance on glass-slide microarraysGenome Biology, 2003
- A Simple Semisolid Subtraction Method Using Carbodiimide-Coated MicroplatesMolecular Biotechnology, 2000
- The solution structure of DNA decamer duplex containing the Dewar product of thymidylyl(3′→5′)thymidine by NMR and full relaxation matrix refinementFEBS Letters, 1998
- Steric factors influencing hybridisation of nucleic acids to oligonucleotide arraysNucleic Acids Research, 1997
- Quantitative Monitoring of Gene Expression Patterns with a Complementary DNA MicroarrayScience, 1995
- Direct fluorescence analysis of genetic polymorphisms by hybridization with oligonucleotide arrays on glass supportsNucleic Acids Research, 1994
- Light-Directed, Spatially Addressable Parallel Chemical SynthesisScience, 1991
- Investigation of nucleic acid secondary structure by means of chemical modification with a carbodiimide reagent. I. Reaction between N-cyclohexyl-N'-β-(4-methylmorpholinium)ethylcarbodiimide and model nucleotidesBiochemistry, 1969