Development of the Handy Bio-Strand and its application to genotyping of OPRM1 (A118G)
2007 (English)In: Analytical Biochemistry, ISSN 0003-2697, E-ISSN 1096-0309, Vol. 367, no 1, 79-86 p.Article in journal (Refereed) Published
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) genotyping of OPRM1 (A118G) by allele-specific oligonucleotide competitive 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 genotyping for small laboratories and hospitals.
Place, publisher, year, edition, pages
2007. Vol. 367, no 1, 79-86 p.
SNP, OPRM1, CYP2CI9, Allele-specific oligonucleotides, Competitive hybridization
IdentifiersURN: urn:nbn:se:uu:diva-103330DOI: 10.1016/j.ab.2007.04.052ISI: 000247868800009PubMedID: 17570330OAI: oai:DiVA.org:uu-103330DiVA: diva2:218060