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Development of the Handy Bio-Strand and its application to genotyping of OPRM1 (A118G)

Ginya, Harumi (author)
Asahina, Junko (author)
Yoshida, Mamiko (author)
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Segawa, Osamu (author)
Asano, Tsutomu (author)
Ikeda, Hideo (author)
Hatano, Yukiko Miyashita (author)
Shishido, Madoka (author)
Johansson, Britt-Marie (author)
Uppsala universitet,Institutionen för farmaceutisk biovetenskap
Zhou, Qin (author)
Uppsala universitet,Institutionen för farmaceutisk biovetenskap
Hallberg, Mathias (author)
Uppsala universitet,Institutionen för farmaceutisk biovetenskap
Takahashi, Masaaki (author)
Nyberg, Fred (author)
Uppsala universitet,Institutionen för farmaceutisk biovetenskap
Tajima, Hideji (author)
Yohda, Masafumi (author)
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 (creator_code:org_t)
Elsevier BV, 2007
2007
English.
In: Analytical Biochemistry. - : Elsevier BV. - 0003-2697 .- 1096-0309. ; 367:1, s. 79-86
Related links:
https://urn.kb.se/re...
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https://doi.org/10.1...
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  • Journal article (peer-reviewed)
Abstract Subject headings
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  • 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.

Subject headings

MEDICIN OCH HÄLSOVETENSKAP  -- Medicinska och farmaceutiska grundvetenskaper -- Farmaceutiska vetenskaper (hsv//swe)
MEDICAL AND HEALTH SCIENCES  -- Basic Medicine -- Pharmaceutical Sciences (hsv//eng)

Keyword

SNP
OPRM1
CYP2CI9
Allele-specific oligonucleotides
Competitive hybridization
PHARMACY
FARMACI

Publication and Content Type

ref (subject category)
art (subject category)

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