Development of the Handy Bio-Strand and its application to genotyping of OPRM1 (A118G).
2007
We previously developed a three-dimensional microarray system, the
Bio-Strand, which exhibits advantages in automated DNA analysis in combination with our Magtration Technology. In the current study, we have developed a compact system for the
Bio-Strand, the Handy
Bio-Strand, which consists of several tools for the preparation of
Bio-Strand Tip, hybridization, and detection. Using the Handy
Bio-Strand, we performed single nucleotide polymorphism (
SNP)
genotypingof OPRM1 (A118G) by
allele-specific oligonucleotidecompetitive hybridization (ASOCH). DNA fragments containing SNP sites were amplified from genomic DNA by PCR and then were fixed on a microporous nylon thread. Thus, prepared
Bio-Strand Tip was hybridized with allele-specific Cy5 probes (<15mer), on which the SNP site was designed to be located in the center. By optimizing the amount of competitors, the selectivity of Cy5 probes increased without a drastic signal decrease. OPRM1 (A118G) genotypes of 23 human genomes prepared from whole blood samples were determined by ASOCH using the Handy
Bio-Strand. The results were perfectly consistent with those determined by PCR direct sequencing. ASOCH using the Handy
Bio-Strand would be a very simple and reliable method for
SNP genotypingfor small laboratories and hospitals.
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