• Journal of Animal Science and Technology
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• v.46 no.2
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• pp.129-136
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• 2004

The objective of present study was to ascertain parentage of Thoroughbred(TB) horses in Korea. A total of 2,029 TB horse samples including 993 foal samples for parentage testing were genotyped for nine international minimum standard markers(AHT4, 5, ASB2, HMS3, 6, 7, HTG4, 10, and VHL20). This method consisted of multiplexing PCR procedure, and showed reasonable amplification of all PCR products. Genotyping was performed with an ABI 310 genetic analyzer. The number of alleles per locus varied from 5 to 11 with a mean value of 7.33 in TB. Expected heterozygosity was ranged from 0.544 to 0.837(mean 0.709) and the total exclusion probability of 9 microsatellites loci was 0.9978. Of the 9 markers, ASB2, HMS7 and HTG10 loci have relatively high PIC value(>0.7). All of the 993 foals were qualified by compatibility according to Mendelian fashion in the present DNA typing for parentage testing. These results suggest that the present DNA typing has high potential for parentage verification of TB horses.

• Asian-Australasian Journal of Animal Sciences
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• v.23 no.4
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• pp.425-429
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• 2010

To develop an efficient DNA typing system for Chinese Holstein cattle, 17 microsatellites, which were amplified in four fluorescent multiplex reactions and genotyped by two capillary electrophoresis injections, were evaluated for parentage verification and identity test. These markers were highly polymorphic with a mean of 8.35 alleles per locus and an average expected heterozygosity of 0.711 in 371 individuals. Parentage exclusion probability with only one sampled parent was approximately 0.999. Parentage exclusion probability when another parent' genotype was known was over 0.99999. Overall probability of identity, i.e. the probability that two animals share a common genotype by chance, was $1.52{\times}10^{-16}$. In a test case of parentage assignment, the 17 loci assigned 31 out of 33 cows to the pedigree sires with 95% confidence, while 2 cows were excluded from the paternity relationship with candidate sires. The results demonstrated the high efficacy of the 17 markers in parentage analysis and individual identification for Chinese Holstein cattle.

• Asian-Australasian Journal of Animal Sciences
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• v.17 no.6
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• pp.750-754
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• 2004

This study was performed for parentage verification of the Korean native horse (KNH). 103 random KNH samples (including 19 foals for parentage testing) were genotyped by using 16 microsatellite markers. The number of alleles per locus varied from 5 to 13 with an average value of 8.56 in the KNH. The observed heterozygosity and the expected heterozygosity ranged 0.398-0.893 (the average value was 0.683) and 0.368-0.871 (the average value was 0.727) in the KNH, respectively. The PIC value and the exclusion probability ranged 0.347-0.853 (the average value was 0.692) and 0.208-0.736, respectively, and the total exclusion probability of 16 microsatellite loci was 0.9999. Of the 16 markers, AHT4, AHT5, ASB2, ASB17, HMS2, HMS3, HTG10, LEX33, TKY321 and VHL20 loci have a relatively high PIC value (>0.7) in the KNH. Of the 19 foals, 5 foals were disqualified by an incompatibility of 4-7 markers according to a Mendelian fashion in the present DNA typing for parentage testing. These results present basic information for developing a system for parentage verification and individual identification in the KNH.

• Journal of Life Science
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• v.15 no.1 s.68
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• pp.45-48
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• 2005

The aim of this study was to construct a correct pedigree of miniature horses (MH). The sex of MH was detected by PCR amplification of the sex determining region of the Y chromosome gene (SRY) prior to parentage testing. Ten random MH samples for parentage testing were genotyped by using 16 micro satellite markers. Since the SRY band (430 bp) was detected in horses No.1, 2, 6, 7, 8, 9, 10, these are male. However, the DNA segment was not identified in horses No.3, 4, and 5, which therefore are female. After genotyping, parentage testing was performed according to Mendelian fashion and International Society for Animal Genetics (ISAG) guideline. Of the 10 MH, 3 were qualified by the compatibility of 16 markers according to Mendelian fashion in the present DNA typing for parentage verification. These results can provide basic information for developing parentage verification and an individual identification system in MH.

• Journal of Embryo Transfer
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• v.28 no.3
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• pp.207-213
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• 2013

This study was carried out to examine a molecular marker system for parentage test in Jeju Black cattle (JBC). Based on the preliminarily studies, we finally selected for construction of a novel genetic marker system for molecular traceability, identity test, breed certification, and parentage test in JBC and its related industrial populations. The genetic marker system had eight MS markers, five indel markers, and two single nucleotide polymorphisms (SNPs; g.G299T and g.del310G) within MC1R gene which is critical to verify the breed specific genotypes for coat color of JBC differing from those of exotic black cattle breeds such as Holstein and Angus. The results showed lower level of a combined non-exclusion probability for second parent (NE-P2) of $4.1202{\times}10^{-4}$ than those previously recommended by International Society of Animal Genetics (ISAG) of $5.000{\times}10^{-4}$ for parentage, and a combined non-exclusion probability for sib identity (NE-SI) of $2.679{\times}10^{-5}$. Parentage analysis has been successfully identified the JBC offspring in the indigenous population and cattle farms used the certified AI semens for production using the JBC-derived offspring for commercial beef. This combined molecular marker system will be helpful to supply genetic information for parentage test and traceability and to develop the molecular breeding system for improvement of animal productivity in JBC population.

• Korean Journal of Veterinary Research
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• v.47 no.4
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• pp.457-460
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• 2007

The objective of the study was to establish routine parentage testing system in Beagle dogs using 22 ISAG (International Society for Animal Genetics) canine microsatellite markers (2005). Blood collections were obtained from a mother dog, 4 candidate father dogs and 3 offspring (n = 8). Genomic DNA samples were extracted from 8 Beagle dogs blood for PCR analysis. PCR products for the allele were analyzed by ABI 3130 DNA Sequencer and GeneScan (Ver 3.0) analysis and Genotyper (Ver. 2.1) software. The genetic relationship of mother and 3 offspring as well as one father dog among 4 candidate father dogs was confirmed by microsatellite allele analysis. The results of locus for amelogenin, which was designed for sexing, were matching with real gender among 8 Beagle dogs (female; 217/217 homozygosity, male; 179/217 heterozygosity). Twenty two ISAG microsatellite markers are useful the parentage test of Beagle dogs. In addition, amelogenin is an applicable marker to detecting real sex in dogs.

• Journal of Life Science
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• v.13 no.4
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• pp.416-420
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• 2003

We performed this study to determine the parentage verification of putative dogs. A total of 7 samples (4 Labrador Retriever dog and 3 Poongsan dog) were genotyped by using 12 international markers (PEZ1, PEZ5, PEZ8, PEZ10, PEZ11, PEZ12, PEZ17, PEZ20, PEZ21, FHC2010, FHC2054, FHC2079). This methods consisted of multiplexing PCR procedures, and it showed reasonable amplification of all PCR products. Genotyping was performed with an ABI 310 genetic analyzer. Labrador Retriever Pup I and Pup II were included according to principles of Mendelian genetics in all loci, while Poongsan Pup III was excluded with markers PEZ1 (106/118), PEZ10 (276/300), and FHC2010 (228/232). These results suggest that the present DNA typing is so useful for parentage verification of these two breeds.

• Horticultural Science & Technology
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• v.34 no.3
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• pp.453-460
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• 2016

The parentage of the horticulturally important pear cultivar 'Niitaka' was confirmed by determining its S-genotypes based on the S-RNase and $PpSFBB^{-{\gamma}}$ genes, and genotyping using simple sequence repeat (SSR) markers. Previous reports suggested that the cultivars 'Amanogawa' and 'Imamuraaki' were the parents of 'Niitaka', although the cultivars 'Chojuro' and 'Shinchu' were also examined as candidate parents, along with two other cultivars. In the present study, the S-genotype of 'Niitaka' was determined to be $S^3S^9$. The $S^9$-RNase of 'Niitaka' was found to be likely inherited from the parent 'Amanogawa' ($S^1S^9$) and the $S^3$-RNase from 'Chojuro' ($S^3S^5$) or 'Shinchu' ($S^3S^5$). Based on the S-genotypes, the cultivar 'Imamuraaki' ($S^1S^6$) had no contribution to the parentage of 'Niitaka' ($S^3S^9$). A total of 67 polymorphic SSR markers were used to further confirm the parentage of 'Niitaka'. Discrepancies were found at several SSR loci between 'Niitaka' and the cultivars 'Imamuraaki' and 'Shinchu', whereas 'Niitaka' inherited alleles from 'Amanogawa' and 'Chojuro' at all SSR loci. Therefore, our findings established that 'Amanogawa' and 'Chojuro' are the parents of pear cultivar 'Niitaka', and not 'Imamuraaki' as previously reported.

• Animal Bioscience
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• v.37 no.4
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• pp.600-608
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• 2024

Objective: In this study, we aimed to evaluate the usability single nucleotide polymorphisms (SNPs) for parentage testing of horse breeds in Korea. Methods: The genotypes of 93 horse samples (38 Thoroughbred horses, 17 Jeju horses, 20 Quarter horses, and 18 American miniature horses) were determined using 15 microsatellite (Ms) markers (AHT4, AHT5, ASB2, ASB17, ASB23, CA425, HMS1, HMS2, HMS3, HMS6, HMS7, HTG4, HTG10, LEX3, and VHL20) and 101 SNP markers. Results: Paternity tests were performed using 15 Ms markers and 101 SNP markers in Thoroughbred horses and Quarter horses. AHT5, ASB2, ASB17, ASB23, CA425, HMS7, HTG10, and LEX3 did not follow Mendelian inheritance in Thoroughbred horses, whereas in Quarter horses, only AHT4, ASB2, and HMS2 showed Mendelian inheritance, consequently, paternity was not established. Meanwhile, 31 markers, including MNEc_2_2_2_98568918_BIEC2_502451, in Thoroughbred horses, and 30 markers, including MNEc_2_30_7430735_BIEC2_816793, in Quarter horses did not conform with Mendelian inheritance and therefore, could not be used for establishing parentage. Conclusion: The possibility of replacing Ms markers with SNP markers for paternity testing in horses was confirmed. However, further research using more samples is necessary.

• Asian-Australasian Journal of Animal Sciences
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• v.22 no.10
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• pp.1351-1358
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• 2009

Parentage tests using polymorphic DNA marker are commonly performed to avoid incorrect recording of the parental information of livestock animals, and single-nucleotide polymorphisms (SNPs) are becoming the method of choice. In Japanese Black cattle, parentage tests based on the exclusion method using microsatellite markers are currently conducted; however, an alternative SNP system aimed at parentage tests has recently been developed. In the present study, two types of simulations were conducted using the pedigree data of two subpopulations in the breed (subpopulations of Hyogo and Shimane prefectures) in order to examine the effect of actual genetic and breeding structures. The first simulation (simulation 1) investigated the usefulness of SNPs for excluding a close relative of the true sire; the second one (simulation 2) investigated the accuracy of sire identification tests for multiple full-sib putative sires by a combined method of exclusion and paternity assignment based on the LOD score. The success rates of excluding a single fullsib and sire of the true sires were, respectively, 0.9915 and 0.9852 in Hyogo and 0.9848 and 0.9852 in Shimane, when 50 SNPs with minor allele frequency (MAF: q) of 0.25${\leq}$q${\leq}$0.35 were used in simulation 1. The success rates of sire identification tests based solely on the exclusion method were relatively low in simulation 2. However, assuming that 50 SNPs with MAF of 0.25${\leq}$q${\leq}$0.35 or 0.45${\leq}$q${\leq}$0.5 were available, the total success rates including achievements due to paternity assignment were, respectively, 0.9430 and 0.9681 in Hyogo and 0.8999 and 0.9399 for Shimane, even when each true sire was assumed to compete with 50 full-sibs.