Validation of StockMarks (R) Set for identifying orgin of species from the canine family

摘要

The goal of this study was to measure the utility of 10 pairs of starter sequences described scientifically to amplify microsatellites in the genome of canines and measure the usefulness of the commercial set StockMarks (R) for Dogs Canine Genotyping Kit from Applied Biosystems company in individual identification. An additional goal is to check the feasibility of using the set to control the origin of species from Canidae familly. The tests were conducted on animals from three species: Red Fox (Vulpes vulpes), Arctic Fox (Alopex lagopus), and Raccoon Dog (Nyctereutes procyonoides), from which blood was drawn into sterile vacuum test-tubes with the anticoagulant EDTA from the saphena vein. DNA was isolated from the blood using QIAamp DNA Blood Mini Kit, a set for isolating genetic material. The obtained products PCR were exposed to electrophoretic separation in the genetic analyzer ABI Prism 3100 Avant, then analyzed using the computer program Gene Mapper (TM) v. 3.5. The frequency of alleles was measured for all microsatellite loci. All calculations were performed using the statistical packet SAS - module SAS/Genetics (TM) 9.1.3.The analysis of the microsatellite sequences (FHC 2010, FHC 2054, FHC 2079, PEZI, PEZ20, PEZ12, PEZ3, PEZ5, PEZ6, PEZ8) in the population of the Arctic Fox, Red fox, and Raccoon Dog, indicates a high degree of polymorphism and high usefulness in the majority of tested microsatellite loci to control the origin of species and identification of specimen in fox and raccoon dog farms. Moreover, it has been confirmed that in many cases clear differences between the length of specific alleles indicates a potential use in identifying species.