Volume 41, Issue 6 (November 1996)

    Rapid and Clear Detection of ABO Genotypes by Simultaneous PCR-RFLP Method

    (Received 17 November 1995; accepted 26 February 1996)

    CODEN: JFSOAD

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    Abstract

    We reported a new approach of ABO genotyping by a polymerase chain reaction and restriction fragment length polymorphism method. Instead of amplifying the loci containing the positions of nucleotides 258 and 700 of cDNA of the A transferase separately, we successfully amplified these 2 loci together in one reaction mixture using 2 sets of primers. The amplified DNA products were digested at the same time with restriction enzymes Kpn I and Alu I. The digested DNA products were then separated by electrophoresis on polyacrylamide gel. In addition, we evaluated the influence of various amplification parameters (concentration of template DNA, primers, Taq DNA polymerase, MgCl2, and number of cycles). In particular, high Mg2+ concentration (3.5 mM) made effective amplification of this locus without producing any unspecific band. By using that optimized condition for PCR, together with a simultaneous approach, our study proved to be time saving, more economic, and convenient in interpreting the results.


    Author Information:

    Wakasugi, C
    Professor, Osaka University Medical School, Osaka,

    Bai, H
    Postgraduate student, Osaka University Medical School, Osaka,

    Kuroki, H
    Research assistant, Osaka University Medical School, Osaka,

    Honda, K
    Associate professor, Osaka University Medical School, Osaka,

    Nakatome, M
    Dental surgeon, Osaka University Medical School, Osaka,

    Nasimul Islam, M
    Postgraduate student, Osaka University Medical School, Osaka,

    Terada, M
    Forensic toxicologist, Osaka University Medical School, Osaka,

    Yamazaki, M
    Research assistant, Osaka University Medical School, Osaka,

    Ogura, Y
    Forensic technician, Osaka University Medical School, Osaka,

    Tun, Z
    Postgraduate student, Osaka University Medical School, Osaka,


    Stock #: JFS14042J

    ISSN: 0022-1198

    DOI: 10.1520/JFS14042J

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    Author
    Title Rapid and Clear Detection of ABO Genotypes by Simultaneous PCR-RFLP Method
    Symposium , 0000-00-00
    Committee E30