One-step immobilization of poly(dT)-modified DNA onto non-modified plastic substrates by UV irradiation for microarrays.
Previously, 'DNattach', an alternative DNA immobilization system for attaching modified oligonucleotide probes onto a gold surface by UV irradiation that can be used in various DNA microarray applications including gene expression analysis, was developed. Attached to the gold surface, the modified probes have been shown to successfully detect synaptogenesis in the developing mouse cerebellum. In this study, this technology to immobilize modified oligonucleotide probes onto three different non-modified plastic surfaces in a microarray format has been further expanded. Using this system, single nucleotide polymorphism (SNP) genotyping of both oligonucleotide and PCR product targets has been successfully performed and it has also been shown that the probes immobilized on the slides can be used efficiently in hybridization experiments. Furthermore, it has been shown that probe concentrations of only 1-5 microM are sufficient for hybridization and that this immobilization method provides hybridization signals greater than those of conventional immobilization techniques.
Kimura
Naoki
N
Research and Development Center, Nisshinbo Industries Inc., 1-2-3 Onodai, Chiba-shi, Chiba 267-0056, Japan. n-kimu@nisshinbo.co.jp
D009693:Nucleic Acid Hybridization / Q000379:methods; D020411:Oligonucleotide Array Sequence Analysis / Q000295:instrumentation / Q000379:methods*; D015345:Oligonucleotide Probes / Q000737:chemistry* / Q000235:genetics / Q000528:radiation effects; D010969:Plastics / Q000737:chemistry*; D017354:Point Mutation / Q000235:genetics; D011071:Poly T / Q000737:chemistry*; D020641:Polymorphism, Single Nucleotide / Q000235:genetics; D015203:Reproducibility of Results; D013499:Surface Properties; D014466:Ultraviolet Rays
D015345:Oligonucleotide Probes; D010969:Plastics; D011071:Poly T