• Biomedical Science Letters
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• v.28 no.4
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• pp.298-306
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• 2022

Pathological tissue fixation using formalin has been widely used for histological samples in many hospitals and institutions. In general, formalin fixatives were either manufactured in laboratories or purchased commercially because of the risks and environmental concerns of handling organic compounds. In this study, the efficacy of three kinds of commercially purchased and one laboratory-made formalin fixative was compared in the PCR-based molecular diagnosis using the extracted DNA from formalin-fixed paraffin-embedded (FFPE) tissues. The quality of extracted DNA from FFPE tonsil tissues with four kinds of formalin solutions was evaluated, and PCR for beta-globin gene and microsatellite instabilities (MSI) tests for pentaplex panel markers were performed using the extracted DNA. There was no difference in PCR and MSI tests as molecular diagnoses regardless of the types of formalin used in this study. However, the total amount and average length of double-stranded DNA extracted from FFPE tonsil tissue showed significant differences according to the type of formalin fixative. Optimized formalin fixatives and methods for DNA extraction might be sophisticated to extract good quality DNA from the small size of specific tissue samples. Further studies are needed to select the most effective formalin fixative for histology and molecular pathology using human FFPE tissues.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.47 no.1
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• pp.52-58
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• 2014

Morphological and genetic variation of two populations of Platichthys stellatus were investigated based on 30 individuals each, collected from Uljin (seedling release area) and Pohang (control) in Korea. Morphological analyses demonstrated that the two populations of P. stellatus were well distinguishable in body color of the blind side and fin shape. Mitochondrial DNA control region analysis indicated no significant differences between the two populations ($F_{ST}=-0.00849$, P>0.05). We also analyzed microsatellite DNA loci of the two populations using six markers. Observed heterozygosity ($H_O$) and expected heterozygosity ($H_E$) were 0.550 and 0.592, respectively, in P. stellatus from Uljin, but 0.700 and 0.737 in P. stellatus from Pohang. An index of differentiation in genetic structure revealed significant differences between the two populations ($F_{ST}=0.0208$, P<0.05). Our results suggest that the Uljin population may be comprised of released P. stellatus, whereas the Pohang population may be wild P. stellatus, highlighting the necessity of continuous monitoring of the two populations.

• Journal of Animal Science and Technology
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• v.45 no.2
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• pp.191-198
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• 2003

This study was carried out to investigate a usefulness of the microsatellite DNA markers for individual identification and parentage verification in three dog breeds. A total of 59 random dog (31 Chiwawa, 20 Poongsan, 8 Labrador Retriever) samples were genotyped by using 14 markers (Chiwawa dog), 16 markers (Poongsan dog), and 12 markers (Labrador Retriever dog) among the 17 international standard markers (PEZ1, 3, 5, 6, 8, 10, 11, 12, 13, 15, 16, 17, 20, 21, FHC2010, FHC2054 and FHC2079), respectively. The number of alleles per locus varied from 4 to 14 with a mean value of 6.07 in Chiwawa dog, 2 to 9 with a mean of 4.75 in Poongsan dog, and 3 to 5 with a mean of 4.00 in Labrador Retriever dog. Observed heterozygosity was ranged 0.419${\sim}$0.968 (mean 0.755), 0.300${\sim}$0.950 (mean 0.597) and 0.125${\sim}$0.750 (mean 0.604), and expected heterozygosity was ranged 0.432${\sim}$0.883 (mean 0.711), 0.262${\sim}$0.817 (mean 0.559) and 0.425${\sim}$0.808 (mean 0.660) in these three dog breeds. PIC value was ranged 0.397${\sim}$0.856 (mean 0.659), 0.222${\sim}$0.772 (mean 0.503) and 0.354${\sim}$0.717 (mean 0.563) in these three dog breeds. Of the 17 markers, PEZ1, PEZ3, PEZ6, PEZ10, PEZ12 loci, PEZ1, PEZ6, PEZ13 loci, and PEZ8, PEZ12 loci have relatively high PIC value (＞0.7) in Chiwawa dog, Poongsan dog and Labrador Retriever dog, respectively. The exclusion probability was ranged 0.240${\sim}$0.741, 0.111${\sim}$0.616, and 0.198${\sim}$0.529, and the combination of microsatellite loci was 0.9999, 0.9991, and 0.9968 in Chiwawa dog, Poongsan dog and Labrador Retriever dog, respectively. These results can give basic information for developing parentage verification and individual identification system in these three dog breeds.

• Food Science of Animal Resources
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• v.27 no.3
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• pp.357-362
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• 2007

Nine brand populations of Hanwoo cattle were characterized using 11 microsatellite DNA markers. The studied populations were: Ansung, Yangpyang, DaeGwanryeng, Palkongsangkangwoo, Hoengseong, Jangsu, Sumjinkang, Hadong, Nam-hae. The observed heterozygosity, expected heterozygosity, and polymorphism information content were calculated. Allele frequencies were calculated and used for the characterization of each brand population and to study their genetic relationships. Genetic distances were estimated using Nei's DA genetic distance and the resultant DA matrix was used in the construction of phylogenetic trees. The NJ tree showed that Ansung and Yangpyang, Sumjinkang and Jangsu, Namhae and Ha-Dong are closely related and are considered to have undergone genetic exchange within the same locale. This study will contribute to the local Hanwoo brand industry.

• Asian-Australasian Journal of Animal Sciences
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• v.21 no.6
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• pp.776-783
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• 2008

In this study, we investigated the genetic diversity and phylogenetic relationship of six duck populations by employing the genetic polymorphisms of 20 microsatellites. The parameters used in this study included number of alleles, average effective numbers of alleles (E) and average rates of heterozygosity of each population. The results showed that all the microsatellite loci were highly polymorphic except that the locus AJ515896 in Muscovy duck was 0. The average PIC (0.762), average h (0.7843) and average E (5.261) of the six duck populations were all high, indicating that the gene polymorphisms and genetic diversity were high. The test of Hardy-Weinberg equilibrium showed that the six populations in this study were all in Hardy-Weinberg disequilibrium. The F-statistic analysis results showed the range of FST was from 0.0205 (AJ515895) to 0.2558 (AJ515896). The mean FST was 0.0936. Phylogenetic study revealed that Peking duck (Z1 and Z4), Shaoxing duck, Cherry Valley duck and Aobaixing duck were clustered in one group, while the Muscovy duck was clustered in one group alone. The phylogenetic relationships among different populations were in accordance with their breeding history and distribution. Our data suggested that the 20 microsatellite loci were effective markers for analysis of genetic relationships among duck populations.

• Journal of Korean Society of Forest Science
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• v.98 no.5
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• pp.568-575
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• 2009

Korean wild and forest cultivated ginseng has long been accepted as high medicinal values compared to field cultivated ginseng. Owing to the high price of Korean wild ginseng, Chinese wild and forest cultivated ginseng were smuggled and sold as Korean wild and forest cultivated ginseng. Therefore, an efficient method is required to distinguish Korean ginseng from Chinese ginseng. Microsatellites, simple sequence repeats (SSRs), are highly polymorphic loci present in DNA that consist of repeating units of base pairs. Thus SSR markers are highly advantageous for detection of small genetic variances of intra-species. In the present study, we constructed a microsatellite-enriched genomic library from South Korean wild Panax ginseng. After sequence analysis of 992 randomly picked positive colonies, 126 (12.7%) of the colonies were found to contain microsatellite sequences, and 38 primer pairs were designed. By polymorphism assessment using 36 primer pairs, 4 primers (PG409, PG450, PG491, and PG582) were shown to be polymorphic to distinguish the South Korean ginseng from the Chinese ginseng. These 4 microsatellite markers will provide powerful tools to authenticate South Korean ginseng from Chinese ginseng.

• Korean journal of applied entomology
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• v.43 no.4 s.137
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• pp.311-315
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• 2004

The brown planthopper, Nilaparvata lugens, is among the most serious insect pests of rice. It is widely distributed in Asia, Australia and Pacific islands. An earlier mitochondrial DNA study revealed that there exist significant genetic differences between populations north and south of the Red River Delta region in Vietnam. However the mitochondrial DNA was not sufficiently variable to examine the sources of immigration. For a more detailed analysis of geographic population structure of N. lugens, we developed microsatellite markers. Thirty-seven putative microsatellite loci were isolated using a magnetic biotin method, and five primer pairs designed from the flanking regions of sequenced microsatellite clones were labeled with fluorescent. Of these five primer sets, two have proven to be useful across all the samples we used in this study. We used variation at these two microsatellite loci to test the hypothesis that N. lugens biotypes (1, 2, and 3) sampled from laboratory selection constituted distinct genetic units. Allele frequency differences among the three major biotype categories were not significantly different at one locus (27035). However, the other (7314) did show differences among the major three biotypes. The methods we describe here will be useful for studying population structure of crop pest and for tracking the patterns of migratory pest like the rice planthoppers.

• Journal of Animal Science and Technology
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• v.51 no.4
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• pp.273-282
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• 2009

DNA marker information is used to identify or distinguish cattle breeds or individual animal. The purpose of this study was to apply Bovine Genotypes Kit Version 1.1/2.1 to bovine DNA samples (National Institute of Animal Science) taken from Australian / American beef (n=148), Holstein beef (n=170) and Hanwoo cattle (n=177) bred in Jeongeub, Jeonbuk, Korea, so that it could distinguish Hanwoo breed. The Bovine Genotype Kits consist of 16 ISAG MS markers, which were used to build a database of genotypes in each group. Genotyping results were analyzed using MS Tool kit and Phylip program to create phylogenetic tree. The GeneClass 2.0 was used to estimate breed identification. These analyses found that this kit had 100% capacity to distinguish Hanwoo beef, 95.3% capacity to differentiate Australian / American beef and 90% capacity to identify Korean Holstein steer beef. Hence, it is expected that 16 commercial microsatellite markers is useful to categorizegenetic characteristics of Hanwoo breed and also identify Hanwoo individuals and the origin of beef. In particular, it is expected that these markers will be advantageous in discriminating domestic Holstein beef from Australian / Americanbeef.

• Journal of Life Science
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• v.21 no.3
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• pp.385-392
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• 2011

The otter (Lutra lutra) in Korea is classified as a first grade endangered species and is managed under state control. We performed a phylogenetic analysis of the otter that inhabits the Changnyeong, Jinju, and Geoje areas in Gyeongsangnamdo, Korea using mtDNA and microsatellite (MS) markers. As a result of the analysis using the 676-bp D-loop sequence of mtDNA, six haplotypes were estimated from five single nucleotide polymorphisms. The genetic distance between the Jinju and Geoje areas was greater than distances within the areas, and the distance between Jinju and Geoje was especially clear. From the phylogenetic tree estimated using the Bayesian Markov chain Monte Carlo analysis by the MrBays program, two subgroups, one containing samples from Jinju and the other containing samples from the Changnyeong and Geoje areas were clearly identified. The result of a parsimonious median-joining network analysis also showed two clear subgroups, supporting the result of the phylogenetic analysis. On the other hand, in the consensus tree estimated using the genetic distances estimated from the genotypes of 13 MS markers, there were clear two subgroups, one containing samples from the Jinju, Geoje and Changnyeong areas and the other containing samples from only the Jinju area. The samples were not identically classified into each subgroup defined by mtDNA and MS markers. It could be inferred that the differential classification of samples by the two different marker systems was because of the different characteristics of the marker systems used, that is, the mtDNA was for detecting maternal lineage and the MS markers were for estimating autosomal genetic distances. Nonetheless, the results from the two marker systems showed that there has been a progressive genetic fixation according to the habitats of the otters. Further analyses using not only newly developed MS markers that will possess more analytical power but also the whole mtDNA are needed. Expansion of the phylogenetic analysis using otter samples collected from the major habitats in Korea should be helpful in scientifically and efficiently maintaining and preserving them.

• Asian-Australasian Journal of Animal Sciences
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• v.31 no.2
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• pp.167-179
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• 2018

Objective: The aim of this study was to create a set of microsatellite markers with high polymorphism for the genetic monitoring and genetic structure analysis of local goose populations. Methods: Novel microsatellite markers were isolated from the genomic DNA of white Roman geese using short tandem repeated probes. The DNA segments, including short tandem repeats, were tested for their variability among four populations of geese from the Changhua Animal Propagation Station (CAPS). The selected microsatellite markers could then be used to monitor genetic variability and study the genetic structures of geese from local geese farms. Results: 14 novel microsatellite loci were isolated. In addition to seven known loci, two multiplex sets were constructed for the detection of genetic variations in geese populations. The average of allele number, the effective number of alleles, the observed heterozygosity, the expected heterozygosity, and the polymorphism information content were 11.09, 5.145, 0.499, 0.745, and 0.705, respectively. The results of analysis of molecular variance and principal component analysis indicated a contracting white Roman cluster and a spreading Chinese cluster. In white Roman populations, the CAPS populations were depleted to roughly two clusters when K was set equal to 6 in the Bayesian cluster analysis. The founders of private farm populations had a similar genetic structure. Among the Chinese geese populations, the CAPS populations and private populations represented different clads of the phylogenetic tree and individuals from the private populations had uneven genetic characteristics according to various analyses. Conclusion: Based on this study's analyses, we suggest that the CAPS should institute a proper breeding strategy for white Roman geese to avoid further clustering. In addition, for preservation and stable quality, the Chinese geese in the CAPS and the aforementioned proper breeding scheme should be introduced to geese breeders.