Probe-on-carriers for oligonucleotide microarrays (DNA chips)

Abstract

Oigonucleotide microarrays (DNA chips) are very efficient tools to analyze genotypes of patients or change in gene expressions between two different samples. However, there is no cost effective procedure to manufacture DNA chips. We are developing ‘probe-on-carriers’, immobilized oligonucleotide probes on solid phase to make DNA chips. In this procedure, each oligonucleotide is synthesized on a controlled porous glass carrier as a solid phase, and can be used as a probe for each sequence. This can be substantiated by technology for strictly controlled pore-size of porous glass. In fact, we found the sequence specific hybridization of probe-on-carrier with using porous glass of larger than 50 nm pore diameter.

The probe-on-carriers for wildtype and mutant p53 genes were hybridized with their complementary probes, respectively, but not with another probes. This result clearly demonstrated that the probe-on-carriers could recognize one-nucleotide substitutions of a gene. We found that the fixed probe-on-carriers on a slide glass still showed sequence specific hybridization. Therefore, we conclude that the probe-on-carriers are epoch-making materials for making DNA chip economically.

Keywords::

• Oligonucleotide microarray
• DNA chip
• Oligonucleotide synthesis
• Hybridization
• Porous glass
• Probe-on-carrier
• Single nucleotide polymorphisms