Aim To develop a reagent kit that enables multiplex polymerase
chain reaction (PCR) amplification of 18 short tandem
repeats (STR) and the canine sex-determining Zinc
Finger marker.
Methods Validation studies to determine the robustness
and reliability in forensic DNA typing of this multiplex
assay included sensitivity testing, reproducibility studies,
intra- and inter-locus color balance studies, annealing
temperature and cycle number studies, peak height
ratio determination, characterization of artifacts such as
stutter percentages and dye blobs, mixture analyses, species-
specificity, case type samples analyses and population
studies.
Results The kit robustly amplified domesticated dog samples
and consistently generated full 19-locus profiles from
as little as 125 pg of dog DNA. In addition, wolf DNA samples
could be analyzed with the kit.
Conclusion The kit, which produces robust, reliable, and
reproducible results, will be made available for the forensic
research community after modifications based on this
study’s evaluation to comply with the quality standards expected
for forensic casework