A Systematic and Quantitative Analysis of PCR Template Contamination

    Volume 45, Issue 6 (November 2000)

    ISSN: 0022-1198

    CODEN: JFSOAD

    Page Count: 5


    Urban, C
    Baxter, Hyland-Immuno Division, Biomedical Research Center, Orth/Donau,

    Falkner, FG
    Baxter, Hyland-Immuno Division, Biomedical Research Center, Orth/Donau,

    Dorner, F
    Baxter, Hyland-Immuno Division, Biomedical Research Center, Orth/Donau,

    Gruber, F
    Baxter, Hyland-Immuno Division, Biomedical Research Center, Orth/Donau,

    Kundi, M
    Institute for Environmental Health, University of Vienna, Vienna,

    Hämmerle, T
    Baxter, Hyland-Immuno Division, Biomedical Research Center, Orth/Donau,

    (Received 21 December 1999; accepted 29 December 1999)

    Abstract

    A quantitative and systematic analysis is provided for ubiquitously present template DNA interfering with the quantification of human DNA by PCR. Two sources contributing to DNA background were identified. The first one is interpreted as DNA present in chemicals and on equipment and the second as caused by operator handling. The amounts were equivalent to 2.5 and 8.9 pg per mL of sample, an the estimated frequencies of contamination were 65 and 35%, respectively, resulting in an effective limit of detection of 17.4 pg/mL. Below this level—named effective laboratory background—a result could not be considered as authentic. Knowledge of these parameters is important for laboratories that analyze minute amounts of human DNA by PCR for purposes such as quantification, typing, and sequencing.


    Paper ID: JFS14885J

    DOI: 10.1520/JFS14885J

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    Author
    Title A Systematic and Quantitative Analysis of PCR Template Contamination
    Symposium , 0000-00-00
    Committee E30