In this study the development of a 13-locus multiplex-PCR system fitting the updated demands for paternity testing in Germany is described. For this purpose an existing multiplex PCR system that allows the simultaneous amplification of eight different STR loci together with the sex-specific locus amelogenin (genRES MPX-2, Serac, Germany) was extended. Whereas some of the primers were taken from the underlying multiplex system, suitable primer sequences were chosen for the STR loci D19S433, TPOX, TH01, D16S539, D5S818, D2S1338 and FGA. Primers of loci resulting in potentially overlapping fragment sizes were labelled with the fluorescent dyes 6-FAM, JOE and NED. Reaction conditions, such as annealing temperature, concentrations of primers and polymerase or buffer conditions were optimised to obtain a robust amplification and reproducible genotype analysis for various sample sources. Full DNA profiles from single source samples were reliably typed from template DNA amounts of as low as 120 pg, suggesting a potential use of this system also in forensic casework analysis. With a mean exclusion chance (MEC) of 99.9989% and a power of discrimination (PD) of about 1×1014 (Caucasians), the new multiplex PCR system provides a significant and sensitive system for forensic DNA analysis. On the basis of these studies, a commercial kit system is now provided by Serac (Bad Homburg, Germany, genRES MPX-3).
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The authors wish to thank Birgit Weber and Waltraud Schatz for their excellent technical assistance and Sabine Woods for critically reading the manuscript.
Author information Authors and AffiliationsInstitute of Legal Medicine, University of Erlangen-Nürnberg, Universitätsstrasse 22, 91054 , Erlangen, Germany
J. Schlenk, S. Seidl, P. Betz & T. Lederer
Serac GmbH, Bad Homburg, Germany
G. Braunschweiger
Correspondence to T. Lederer.
About this article Cite this articleSchlenk, J., Seidl, S., Braunschweiger, G. et al. Development of a 13-locus PCR multiplex system for paternity testing. Int J Legal Med 118, 55–61 (2004). https://doi.org/10.1007/s00414-003-0420-5
Received: 01 October 2003
Accepted: 27 November 2003
Published: 14 January 2004
Issue Date: February 2004
DOI: https://doi.org/10.1007/s00414-003-0420-5
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