Journal Published Online: 01 November 2000
Volume 45, Issue 6

A Systematic and Quantitative Analysis of PCR Template Contamination

CODEN: JFSCAS

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
Hyland-Immuno Division, Biomedical Research Center, Orth/Donau, Austria
Gruber, F
Hyland-Immuno Division, Biomedical Research Center, Orth/Donau, Austria
Kundi, M
Institute for Environmental Health, University of Vienna, Vienna, Austria
Falkner, FG
Hyland-Immuno Division, Biomedical Research Center, Orth/Donau, Austria
Dorner, F
Hyland-Immuno Division, Biomedical Research Center, Orth/Donau, Austria
Hämmerle, T
Hyland-Immuno Division, Biomedical Research Center, Orth/Donau, Austria
Pages: 5
Price: $25.00
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Stock #: JFS14885J
ISSN: 0022-1198
DOI: 10.1520/JFS14885J