Original scientific paper
Developmental Validation of Short Tandem Repeat Reagent Kit for Forensic DNA Profiling of Canine Biological Material
Melody Dayton
; The Jan Bashinski DNA Laboratory, California Department of Justice, Richmond, Calif, USA
Mikko T. Koskinen
; Finnzymes Diagnostic, Espoo, Finland
Bradley K. Tom
; Graduate Group in Forensic Science, University of California-Davis, Davis, Calif, USA
Anna-Maria Mattila
; Finnzymes Diagnostic, Espoo, Finland
Eric Johnston
; MMI Genomics, Inc, Davis, Calif, USA
Joy Halverson
; QuestGen Forensics, Davis, Calif, USA
Dennis Fantin
; MMI Genomics, Inc, Davis, Calif, USA
Sue DeNise
; MMI Genomics, Inc, Davis, Calif, USA
Bruce Budowle
; Institute of Investigative Genetics, University of North Texas Health Science Center, Fort Worth, Tex, USA
David Glenn Smith
; Finnzymes Diagnostic, Espoo, Finland
Sree Kanthaswamy
; Department of Anthropology, University of California-Davis, Davis, Calif, USA
Abstract
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.
Keywords
domestic dog; microsatellites; forensic science; multiplex; optimization; non-human; DNA
Hrčak ID:
40680
URI
Publication date:
15.6.2009.
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