Peter Weisner¹, Peder Carstenius¹, David Spodick¹ and Angel Carracedo^2
1 Pharmacia Biotech, Alameda, CA
² Institute of Legal Medicine, Santiago de Compostela, Spain

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On-line detection of fluorescent labeled STR systems using automated DNA sequencers has become a widely accepted method in genetic identification. High throughput can be achieved using the Pharmacia ALF express DNA Analysis System by high speed electrophoresis (1h/gel/40 lanes) and the latest multiplexing systems. However, the limiting bottleneck with all commercially available DNA Sequencers used to be the labour and time consuming user-interactive allele assignment.

Our approach is to use highly standardized temperature controlled electrophoresis conditions, internal standards in every lane and non-overlapping multiplex systems. A new software has been developed for automated allele calling using allelic ladders as external sizing standards. The high intra- and inter- gel sizing reproducibility allows the use of very stringent size windows (typically: +/- 0.5 bases ) which is necessary to avoid errors or ambiguities in fully automated allele calling.

Methodology and data will be presented using Promega Multiplexing systems and the new Pharmacia software for fully automated allele calling of various samples.