Attachment of oligonucleotide probes to poly carbodiimide-coated glass for microarray applications.
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 approximately 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.
Kimura
Naoki
N
Research and Development Center, Nisshinbo Industries Inc., 1-2-3 Ohnodai, Midori-ku, Chiba-shi, Chiba 267-0056, Japan. n-kimu@nisshinbo.co.jp
Oda
Ryuichi
R
Inaki
Yoshiaki
Y
Suzuki
Osamu
O
D010582:Bacteriophage lambda / Q000235:genetics; D002234:Carbodiimides / Q000528:radiation effects; D004279:DNA, Viral / Q000235:genetics; D020869:Gene Expression Profiling / Q000295:instrumentation; D005898:Glass; D006801:Humans; D009693:Nucleic Acid Hybridization; D020411:Oligonucleotide Array Sequence Analysis / Q000295:instrumentation*; D015345:Oligonucleotide Probes / Q000235:genetics / Q000378:metabolism*; D009841:Oligonucleotides / Q000235:genetics / Q000378:metabolism; D016133:Polymerase Chain Reaction; D020641:Polymorphism, Single Nucleotide / Q000235:genetics; D015203:Reproducibility of Results; D014466:Ultraviolet Rays
D002234:Carbodiimides; D004279:DNA, Viral; D015345:Oligonucleotide Probes; D009841:Oligonucleotides