• Reproductive and Developmental Biology
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• v.29 no.4
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• pp.213-221
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• 2005

We used nine decamer primers to generate DNA fragment sizes ranging from 100 bp to 1,600 bp from two bullhead (Pseudobagrus fulvidraco) populations of Dangjin in Korea. 376 fragments were identified in the cultured bullhead population, and 454 in the population of wild bullhead from Dangjin: 287 specific fragments $(76.3\%)$ in the cultured bullhead population and 207 $(45.6\%)$ in the wild bullhead population. On average, a decamer primer was used to generate 34.2 amplified products in a cultured bullhead. A RAPD primer was used to generate an average of 3.1 amplified bands per sample, ranging between 2.5 and 6.0 fragments in this population. Nine primers also generated 24 polymorphic fragments (24/376 fragment, $6.4\%$) in the cultured bullhead population, and 24 (24/454 fragments, $5.2\%$) in the wild bullhead population. The OPA-16 primer, notably, produced which 11 out of 11 bands $(100\%)$ were monomorphic in the wild bullhead population. 110 intra-population-specific fragments, with an average of 12.2 per primer, were observed in the cultured bullhead population. 99 fragments, with an average of 11.0 per primer, were identified in the wild bullhead. Especially, 55 inter-population-common fragments, with an average of 6.1 per primer, were observed in the two bullhead populations. The bandsharing value (BS value) of individuals within the wild bullhead population was substantially higher than was determined in the cultured bullhead population. The average bandsharing value was $0.596\pm0.010$ within the cultured bullhead population,. and $0.657\pm0.010$ within the wild bullhead population. The dendrogram obtained with the nine primers indicates two genetic clusters, designated cluster $1\;(CULTURED\;01\~CULTURED\;11)$, and cluster $2\;(WILD\;12\~WILD\;22)$. Ultimately, the longest genetic distance displaying significant molecular differences was determined to exist between individuals in the two bullhead populations, namely between individuals WILD no. 19 of the wild bullhead population and CULTURED no. 03 of the cultured bullhead population (genetic distance = 0.714). RAPD-PCR allowed us to detect the existence of population discrimination and genetic variation in Korean population of bullhead. This finding indicates that this method constitutes a suitable tool for DNA comparison, both within and between individuals, populations, species, and genera.

• Korean Journal of Ichthyology
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• v.17 no.3
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• pp.173-186
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• 2005

Genomic DNA was isolated from two geographic populations of largehead hairtail (Trichiurus lepturus) in Korea and the Atlantic Ocean. The eight arbitrarily selected primers were found to generate common, polymorphic, and specific fragments. The complexity of the banding patterns varied dramatically between primers from the two locations. The size of the DNA fragments also varied widely, from 150 bp (base pairs) to 3,000 bp. Here, 947 fragments were identified in the largehead hairtail population from Korea, and 642 in the largehead hairtail population from the Atlantic Ocean: 148 specific fragments (15.6%) in the Korean population, and 61 (9.5%) in the Atlantic population. In the Korean population, 638 common fragments with an average of 79.8 per primer were observed.; 429 common fragments, with an average of 53.6 per primer, were identified in the Atlantic population. The number of polymorphic fragments in the largehead hairtail population from Korea and the Atlantic Ocean was 76 and 27, respectively. Based on the average bandsharing values of all samples, the similarity matrix ranged from 0.784 to 0.922 in the Korean population, and from 0.833 to 0.990 in the Atlantic population. The bandsharing value of individuals within the Atlantic population was much higher than in the Korean population. The dendrogram obtained by the eight primers indicated two genetic clusters: cluster 1 (KOREAN 01~KOREAN 11), and cluster 2 (ATLANTIC 12~ATLANTIC 22). Individual KOREAN no. 10 from Korea was genetically most closely related to KOREAN no. 11 in the Korean population (genetic distance = 0.038). Ultimately, individual KOREAN no. 01 of the Korean population was most distantly related to ATLANTIC no. 16 of the Atlantic population (genetic distance = 0.708).

• Development and Reproduction
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• v.20 no.4
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• pp.379-385
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• 2016

Seven oligonucleotides primers were shown to generate the shared loci, specific loci, unique shared loci to each species and shared loci by the three species which could be obviously scored. In the present study, 7 oligonucleotides primers produced 401 total loci in the Styela clava (SC) species, 390 in the Halocynthia roretzi (HR) and 434 in the Styela plicata (SP), respectively. Seven oligonucleotides primers generated 275 specific loci in the SC, 341 in the HR and 364 in the SP species, respectively. The oligonucleotides primer BION-23 generated 28 unique loci to each species in the SP species. Especially, the oligonucleotides primer BION-25 produced 7 unique loci to each species, which were identifying each species in the SP species. BION-17 distinguished 21 shared loci by the three ascidian species, major and/or minor fragments of sizes, which were identical in almost all of the samples. Based on the average bandsharing values of all samples, the similarity matrix ranged from 0.519 to 0.774 in the SC species, from 0.261 to 0.683 in the HR species and from 0.346 to 0.730 in the SP species. As regards average bandsharing value (BS) results, individuals from SC species ($0.661{\pm}0.081$) exhibited higher bandsharing values than did individuals from HR species ($0.555{\pm}0.074$) (P<0.05). The dendrogram obtained by the seven oligonucleotides primers indicates three genetic groups. In three ascidian species, the shortest genetic distance (0.071) exhibiting significant molecular difference was also between individual no. 20 and no. 21 within the SP species.

• Journal of Aquaculture
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• v.17 no.1
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• pp.69-80
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• 2004

The objective of the present study was to analyze genetic distances, variation and characteristics of individuals in rainbow trout, Oncorhynchus mykis using amplified fragment length polymorphism (AFLP) method as molecular genetic technique, to detect AFLP band patterns as genetic markers, and to compare the efficiency of agarosegel electrophoresis (AGE) and polyacrylamide gel electrophoresis (PAGE), respectively. Using 9 primer combinations, a total of 141 AFLP bands were produced, 108 bands (82.4%) of which were polymorphic in AGE. In PAGE, a total of 288 bands were detected, and 220 bands (76.4%) were polymorphic. The AFLP fingerprints of AGE were different from those of PAGE. Separation of the fragments with low molecular weight and genetic polymorphisms revealed a distinct pattern in the two gel systems. In the present study, the average bandsharing values of the individuals between two populations apart from the geographic sites in Kangwon-do ranged from 0.084 to 0.738 of AGE and PAGE. The bandsharing values between individuals No.9 and No. 10 showed the highest level within population, whereas the bandsharing values between individuals No.5 and No.7 showed the lowest level. As calculated by bandsharing analysis, an average of genetic difference (mean$\pm$SD) of individuals was approximately 0.590$\pm$0.125 in this population. In AGE, the single linkage dendrogram resulted from two primers (M11+H11 and M13+H11), indicating six genetic groupings composed of group 1 (No.9 and 10), group 2 (No. 1, 4, 5, 7, 10, 11, 16 and 17), group 3 (No. 2, 3, 6, 8, 12, 15 and 16), group 4 (No.9, 14 and 17), group 5 (No. 13, 19, 20 and 21) and group 6 (No. 23). In AGE, the genetic distances among individuals of between-population ranged from 0.108 to 0.392. In AGE, the shortest genetic distance (0.108) displaying significant molecular differences was between individuals No.9 and No. 10. Especially, the genetic distance between individuals No. 23 and the remnants among individuals within population was highest (0.392). Additionally, in the cluster analysis using the PAGE data, the single linkage dendrogram resulted from two primers (M12+H13 and M11+H13), indicating seven genetic groupings composed of group 1 (No. 15), group 2 (No. 14), group 3 (No. 11 and 12), group 4 (No.5, 6, 7, 8, 10 and 13), group 5 (No.1, 2, 3 and 4), group 6 (No.9) and group 7 (No. 16). By comparison with the individuals in PAGE, genetic distance between No. 10 and No. 7 showed the shortest value (0.071), also between No. 16 and No. 14 showed the highest value (0.242). As with the PAGE analysis, genetic differences were certainly apparent with 13 of 16 individuals showing greater than 80% AFLP-based similarity to their closest neighbor. The three individuals (No. 14, No. 15 and No. 16) of rainbow trout between two populations apart from the geographic sites in Kangwon-do formed distinct genetic distances as compared with other individuals. These results indicated that AFLP markers of this fish could be used as genetic information such as species identification, genetic relationship or analysis of genome structure, and selection aids for genetic improvement of economically important traits in fish species.

• Development and Reproduction
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• v.16 no.4
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• pp.315-320
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• 2012

The gDNA isolated from Cyclina sinensis from Gochang (GOCHANG), Incheon (INCHEON) and a Chinese site (CHINESE), were amplified by PCR. Here, the seven oligonucleotide decamer primers (BION-66, BION-68, BION-72, BION-73, BION-74, BION-76, and BION-80) were used to generate the unique shared loci to each population and shared loci by the three cyclina clam populations. As regards multiple comparisons of average bandsharing value results, cyclina clam population from Chinese (0.763) exhibited higher bandsharing values than did clam from Incheon (0.681). In this study, the dendrogram obtained by the seven decamer primers indicates three genetic clusters: cluster 1 (GOCHANG 01~GOCHANG 07), cluster 2 (INCHEON 08~INCHEON 14), cluster 3 (CHINESE 15~CHINESE 21). The shortest genetic distance that displayed significant molecular differences was between individuals 15 and 17 from the Chinese cyclina clam (0.049), while the longest genetic distance among the twenty-one cyclina clams that displayed significant molecular differences was between individuals GOCHANG no. 03 and INCHEON no. 12 (0.575). Individuals of Incheon cyclina clam population was somewhat closely related to that of Chinese cyclina clam population. In conclusion, our PCR analysis revealed a significant genetic distance among the three cyclina clam populations.

• Development and Reproduction
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• v.22 no.3
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• pp.283-288
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• 2018

The author carried out PCR-based genetic platform to investigate the hierarchical polar dendrogram of Euclidean genetic distances of one bastard halibut population, particularly for Paralichthys olivaceus, which was further connected with those of the other fish population, by involving with the precisely designed oligonucleotide primer sets. Eight oligonucleotides primers were used generating excessively alterating fragments, ranging in size of DNA bands from larger than approximately 100 bp to less than 2,000 bp. As regards average bandsharing value (BS) results, individuals from Hampyeong population (0.810) displayed lower bandsharing values than did individuals from Wando population (0.877). The genetic distance between individuals approved the existence of close relationship in the cluster II. Relatively, individuals of one bastard halibut population were fairly related to that of the other fish population, as shown in the polar hierarchical dendrogram of Euclidean genetic distances. The points of a noteworthy genetic distance between two P. olivaceus populations demonstrated this PCR procedure is one of the quite a few means for individuals and/or populations biological DNA investigates, for species security and proliferation of bastard halibut individuals in coastal region of the Korea.

• The Korean Journal of Malacology
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• v.32 no.4
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• pp.255-261
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• 2016

The selected seven oligonucleotides primers BION-32, BION-33, BION-35, BION-38, BION-40, BION-46 and BION-58 generated the shared loci, specific loci, unique shared loci to each population and shared loci by the three T. granosa populations in Beolgyo, a Chinese site and Wonsan, respectively. The bandsharing value between individuals' no. 03 and no. 04 was 0.816, which was the highest value identified within the Beolgyo population. The primer BION-35 generated the most loci (a total of 70), with an average of 10.0 in the Wonsan population. On average, seven oligonucleotides primers generated 16.1 specific loci in the Beolgyo population, 22.3 in the Chinese population and 39.3 in the Wonsan population. 126 unique shared loci to each population, with an average of 18 per primer, were observed in the Beolgyo population, 63 loci, with an average of 9 per primer, were observed in the Chinese population, and 49 loci, with an average of 7 per primer, and were observed in the Wonsan population. The oligonucleotides primer BION-32 generated 14 unique loci to each population, which were identifying each population in the Beolgyo population. Interestingly, every primer had not distinguished the shared loci by the three populations, major and/or minor fragments of sizes, which were identical in almost all of the samples. As regards average bandsharing value (BS) results, individuals from Beolgyo population ($0.717{\pm}0.057$) exhibited higher BS values than did those from Wonsan population ($0.552{\pm}0.104$) (P < 0.05). The dendrogram resulted from truthful seven oligonucleotides primers, representing three genetic clusters comprising group I (BEOLGYO 01, 02, 03, 04, 05, 06 and 07), group II (CHINESE 08, 09, 10, 11, 12, 13 and 14) and group III (WONSAN 15, 16, 17, 18, 19, 20 and 21). In three T. granosa populations, the longest genetic distance (0.874) displaying significant molecular difference was also between individual no. 02 within the Beolgyo population and individual no. 12 within the Chinese population. Relatively, individuals of the CHINESE population were fairly closely related to those of the WONSAN population.

• Development and Reproduction
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• v.5 no.2
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• pp.151-158
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• 2001

Genomic DNA from the blood of crucian carp(Carassius carassiu) from lake and aquaculture facility in Kunsan, Korea was extracted in order to identify genetic differences by polymerase chain reaction-randomly amplified polymorphic DNAs(PCR-RAPD). Out of 12 primers, 6 generated 266 highly reproducible RAPD markers, producing approximately 2.1 polymorphic bands per primer in crucian carp from lake. The degree of similarity varied from 0.18 to 0.76 as calculated by bandsharing analysis in crucian carp from lake. The RAPD outlines obtained with DNA of two different crucian carp populations from Kunsan were different(0.47 from lake and 0.70 from aquaculture facility, respectively). The electrophoretic analysis of polymerase chain reaction-randomly amplified polymorphic DNAs(PCR-RAPD) products showed high levels of similarity between different individuals in crucian carp from aquaculture facility. This result implies the genetic similarity due to raising in the same environmental condition or inbreeding within the crucian carp from aquaculture facility in Kunsan. In other words, crucian carp may have high levels of genome DNA diversity due to the introduction of the wild population from the other sites of Knsan even if it may be the geographical diverse distribution of this species. Generally, the RAPD polymorphism generated by these primers may be useful as a genetic marker for strain or population identification of important aquacultural fish species, crucian carp. However, in future, additional populations and sampling sites will be necessary to complement weak points.

• Asian-Australasian Journal of Animal Sciences
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• v.17 no.8
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• pp.1053-1061
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• 2004

Of the 20 arbitrarily chosen primers, six oligonucleotides decamer primers were used on the basis of the number of the polymorphisms generated in catfish (Silurus asotus) from Yesan and bullhead (Pseudobagrus fulvidraco) from Dangjin in Korea. Six primers were used generating a total of 602 scorable bands in catfish and 195 in bullhead population, respectively, ranging in size of DNA fragments from less than approximately 100 to larger than 2,000 base pairs (bp). Six primers yielded 199 polymorphic fragments (33.1%) in catfish and 47 (24%) in bullhead, respectively. In the present study, a total of 328 common fragments (an average of 54.7 per primer) were observed in catfish population, whereas 84 (an average of 14.0 per primer) in bullhead. The total number of specific fragments in catfish and bullhead population were 76 and 64, respectively. In catfish population, random decamer, OPA-17 (GACCGCTTGT) generated the highest number of fragments (a total of 141) in comparison with other primers used, with an average of 11.8. The common bands in the molecular weight of 300 bp generated by random primer OPA-06 (GGTCCCTGAC) were present in every individuals in bullhead population. The major polymorphic bands in the molecular weight of 100 bp generated by OPA-17 were identified in lane 14, 15, 17, 18, 19 20 and 21, which were identifying species in bullhead population. The average bandsharing values (BS values) of all of the samples within catfish population ranged from 0.575 to 0.945, whereas 0.063-1.000 within bullhead population. The bandsharing value (index of similarity between individuals) between individual No. 5 and No. 9 showed the highest level within catfish population, whereas the bandsharing value between individual No. 1 and No. 2 showed the lowest level. The single linkage cluster analysis resulted from four primers, indicating four genetic groupings composed of group 1 (C1-C10, all of the catfish samples), group 2 (B11, B12, B13, B14, B16, B17, B18, B19), group 3 (B15) and group 4 (B20 and B21). The dendrogram reveals close relationships between individual identities within two species populations and individuals derived from the same ancestor, respectively. However, genetic distances between two species populations ranged from 0.124 to 0.333. The shortest genetic distance (0.042) displaying significant molecular differences was between individual No. 6 and No. 9 catfish population. The shortest genetic distance (0.033) displaying significant molecular differences also was between individual No. 18 and No. 19 in bullhead population. Reversely, the genetic distance of individual No. 20/21 among individuals in bullhead population was highest (0.333). This result showed that bullhead No. 20 and 21 were distinct from other individuals within bullhead population.

• The Korean Journal of Malacology
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• v.25 no.1
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• pp.5-13
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• 2009

Genomic DNA isolated from two geographical ark shell (Scapharca subcrenata) populations was amplified several times by PCR reactions. The ark shell population from Daecheon (ASPD) and from Wonsan (ASPW) in the West Sea and the East Sea of Korean Peninsula, respectively, obtained. The seven arbitrarily selected primers OPA-05, OPA-11, OPB-09, OPB-11, OPB-14, OPC-18 and OPD-07 were shown to generate the loci observed per primer, shared loci by each population, specific loci, unique shared loci to each population and shared loci by the two populations which could be clearly scored. Here, 862 loci were identified in the ASPD population, and 1,191 in the ASPW population: 137 specific loci (15.9%) in the Daecheon population and 84 (7.1%) in the Wonsan population. 407 shared loci by each population, with an average of 58.1 per primer, were observed in the ASPD population. 473 shared loci by each population, with an average of 67.6 per primer, were identified in the ASPW population. The numbers of specific loci in the ASPD and ASPW population were 137 and 84, respectively. Consequently, the average bandsharing value of individuals within the ASPW population was much higher than in the ASPD population. The bandsharing value between individuals' no. 08 and no. 13 was 0.628, which was the highest measured between the two geographical populations. The dendrogram obtained by the seven primers indicated three genetic clusters: cluster 1 (DAECHEON 01-DAECHEON 11), cluster 2 (WONSAN 12 and 14) and cluster 3 (WON SAN 13, 15, 16, 17, 18, 19, 20, 21 and 22). The genetic distance between the two geographical populations ranged from 0.043 to 0.499. Especially, individual no. 10 of Daecheon population was most distantly related to no. 14 of Wonsan population (genetic distance = 0.499).