Volume 45, Issue 6 (November 2000)

    A Systematic and Quantitative Analysis of PCR Template Contamination

    (Received 21 December 1999; accepted 29 December 1999)

    CODEN: JFSOAD

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    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.


    Author Information:

    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,


    Stock #: JFS14885J

    ISSN: 0022-1198

    DOI: 10.1520/JFS14885J

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