Evaluation of the ALF DNA sequencer for high-speed sizing of short tandem repeat alleles

Electrophoresis
R Decorte, J J Cassiman

Abstract

DNA typing of short tandem repeat (STR) loci with automated real-time analysis of the fluorescent polymerase chain reaction (PCR) products has given forensic DNA analysis a new dimension. In the present work, the ALF DNA sequencer was evaluated for automated size determination of tetra-nucleotide STRs at high speed. Short gel plates, with a well-to-laser distance of 10 cm, allowed for the analysis of four STR loci (HUMvWF, HUMTH01, D21S11 and HPRT) in one gel lane in less than 75 min. Allele size determination was done with two external allelic ladders for each locus. Lane-to-lane variations were overcome by the inclusion in each lane of two fluorescent PCR products of constant size (123 and 375 bp) that migrated below and above the multiplex of the four STR loci. The accuracy of sizing and allele detection within and between different gels was high (99.89%) for all four STR systems investigated and the gels could be reloaded without a decrease in accuracy of the allele size estimation. This way, the throughput of the system was increased, which is of interest for linkage studies, gene mapping, and population diversity studies.

References

Apr 1, 1992·Human Molecular Genetics·V Sharma, M Litt
Aug 1, 1992·PCR Methods and Applications·K M SullivanJ M Robertson
Apr 1, 1992·Genomics·J S Beckman, J L Weber
Jul 1, 1990·DNA and Cell Biology·R DecorteJ J Cassiman
Jan 1, 1994·International Journal of Legal Medicine·A MöllerB Brinkmann
Aug 1, 1993·PCR Methods and Applications·C P KimptonM Adams
Jan 1, 1993·International Journal of Legal Medicine·P WiegandB Brinkmann

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Citations

Jan 17, 2002·Forensic Science International : Synergy·B ZaharovaI Kremensky
Jul 22, 1998·Forensic Science International : Synergy·E JehaesR Decorte
Jan 8, 2000·Arthroscopy : the Journal of Arthroscopic & Related Surgery : Official Publication of the Arthroscopy Association of North America and the International Arthroscopy Association·P DebeerJ Bellemans
Jan 11, 2000·The American Journal of Forensic Medicine and Pathology·D De LeoM Marigo
Feb 25, 2009·The American Journal of Forensic Medicine and Pathology·Aleksandar ApostolovEkaterina Angelova
Mar 4, 2014·Biotechnology, Biotechnological Equipment·Аleksandar Apostolov

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