Development of a 19‐plex short tandem repeat typing system for individual identification and parentage testing of horses (Equus caballus)

Abstract

Summary Individual identification of horses for pedigree verification and registration is important for the sustainable development of the horse industry. Horse individual identification and parentage tests commonly use the 17 short tandem repeats (STRs) recommended by the International Society for Animal Genetics (ISAG) and the locus LEX33. While many multiplex STR typing systems have been established for the horse, a sex determining marker is usually absent, and none of them can simultaneously detect all 17 ISAG recommended loci and the locus LEX33. Here, we present a 19‐plex STR typing system that contains the 17 ISAG recommend loci, the locus LEX33 and amelogenin as sex determining loci. The results of our sensitivity, species specificity, stutter analysis and population data analysis, indicate that this system is a specific, sensitive, and robust tool for the identification of individuals, parentage testing and genetic research in the horse.

Short Tandem Repeat DNA Profiling Using Perylene-Oligonucleotide Fluorescence Assay

“…This property enables one to identify a specific individual based on genotyping the STR variations. This application is the basis of widely used forensic applications, , population and disease studies, − as well as cell line profiling. − Their presence is also related to secondary DNA motifs like hairpins or G quadruplex complexes and serious pathologies such as cancer, , among others. − …”

Changes in population structure and genetic diversity of Misaki horses between 2015 and 2020

“…From the blood samples with added EDTA as anticoagulant, DNA was extracted using the QIAamp DNA Blood Mini Kit (QIAGEN K.K., Tokyo, Japan). A total of 31 microsatellite markers (AHT4, AHT5, ASB2, ASB17, ASB23, CA425, and HMS2, HMS3, HMS6, HMS7, HTG4, HTG10, LEX33, TKY19, TKY28, TKY321, VHL20, TKY279, TKY287, TKY294, TKY297, TKY301, TKY312, TKY325, TKY333, TKY337 341, TKY343, TKY344, TKY374, TKY394) were used [ 9 , 12 ] and were genotyped as previously reported [ 3 , 6 , 7 , 8 , 10 , 11 ]. Allele discrimination was based on the complementation panel adopted at the International Society of Animal Genetics workshop to determine equine parentage [ 9 ].…”

“…A total of 31 microsatellite markers (AHT4, AHT5, ASB2, ASB17, ASB23, CA425, and HMS2, HMS3, HMS6, HMS7, HTG4, HTG10, LEX33, TKY19, TKY28, TKY321, VHL20, TKY279, TKY287, TKY294, TKY297, TKY301, TKY312, TKY325, TKY333, TKY337 341, TKY343, TKY344, TKY374, TKY394) were used [ 9 , 12 ] and were genotyped as previously reported [ 3 , 6 , 7 , 8 , 10 , 11 ]. Allele discrimination was based on the complementation panel adopted at the International Society of Animal Genetics workshop to determine equine parentage [ 9 ]. From these results, the average number of microsatellite alleles ( Na ), expected heterozygosity ( He ), observed value ( Ho ), and fixation index (F is ) were calculated using GENEPOP [ 4 ].…”