Successful direct amplification of nuclear markers from single dog hairs using DogFiler multiplex

We report on successful amplification of canine STR DNA profiles from single dog hairs. Dog hairs are commonly found on clothing or items of interest in forensic casework and may be crucial associative evidence if linked to an individual dog. We used direct amplification from these hairs to increase the DNA yield of the sample, as well as greatly reducing analysis time. Hairs from different somatic regions were used from several different dog breeds to amplify a selection of eight loci from the validated DogFiler multiplex. Naturally shed canine hairs were processed, with a mix of coarse topcoat (guard) hairs and thinner soft undercoat hairs. Multiple sections of single hairs were amplified in 5 mm segments to determine the viability of DNA recovery from the shaft of the hair. Single guard hairs were cut into 5 mm sections and added directly into a PCR tube. Undercoat hairs, which are very fine, were amplified together in a single tube (approximately ten small hairs). Coarse hairs were found to be the most successful in producing full DNA profiles at all eight loci, matching the corresponding reference profile for that dog.