Developmental Validation of Short Tandem Repeat Reagent Kit for Forensic DNA Profiling of Canine Biological Materials

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. ResultsThe 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. ConclusionThe 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.

Design, installation, and performance evaluation of a custom dye matrix standard for automated capillary electrophoresis

“…Automated CE is designed to separate, detect, and analyze fluorescently labeled DNA fragments, and allows for a wide range of applications such as standard PCR fragment analysis , minisequencing , amplified fragment length polymorphism , or Sanger sequencing (reviewed in ). Numerous commercial genotyping kits are available, either for identification of human individuals (e.g., Applied Biosystems' GlobalFiler™ PCR Amplification Kit , Promega's PowerPlex 21 , and Qiagen's Investigator DIPplex kits), domestic animals (e.g., Applied Biosystems' StockMarks ), cultivars (e.g., Labindia's Basmati Verifiler Kit ), or bacterial species (e.g., MseI Microbial AFLP Ligation/Amplification Module ).…”

Field Testing of Collection Cards for Cannabis sativa Samples With a Single Hexanucleotide DNA Marker*,†

“…The development of short tandem repeat (STR) analysis has streamlined human individualization and now allowed for DNA genotyping of many additional species such as cat (17,18), dog (19)(20)(21)(22)(23)(24), horse (25)(26)(27)(28)(29)(30), and marijuana (31)(32)(33)(34)(35)(36)(37) that have significant value as trace evidence. There are several scientific reports about STR markers in development for Cannabis sativa (31)(32)(33)(34)(35)(36)(37).…”

Molecular genetic diversity of donkey (Equus asinus) in South Korea

“…As compared to genotype frequency, gene frequency is very stable beyond the generation, so usually, many studies of molecular evolution deal with changes in gene frequency. However, what is directly related to the phenotype of individual is genotype [7, 20].…”