Developmental validation of short tandem repeat reagent kit for forensic DNA profiling of canine biological material.
OBJECTIVE
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.


CONCLUSIONS
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.
Dayton
Melody
M
Department of Anthropology, University of California, One Shields Avenue, Davis, CA 95616, USA.
Koskinen
Mikko T
MT
Tom
Bradley K
BK
Mattila
Anna-Maria
AM
Johnston
Eric
E
Halverson
Joy
J
Fantin
Dennis
D
DeNise
Sue
S
Budowle
Bruce
B
Smith
David Glenn
DG
Kanthaswamy
Sree
S
D000818:Animals; D004247:DNA / Q000235:genetics*; D016172:DNA Fingerprinting / Q000379:methods*; D004285:Dogs; D053803:Forensic Genetics; D018895:Microsatellite Repeats; D016133:Polymerase Chain Reaction / Q000379:methods; D011933:Reagent Kits, Diagnostic
D011933:Reagent Kits, Diagnostic; D004247:DNA