• Proceedings of the PSK Conference
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• 2003.10b
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• pp.85.1-85.1
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• 2003

Previously, we found the operon random primer (OP-5A) that is characteristic the genus Panax by randomly amplified polymorphic DNA (RAPD) analysis. However, OP-5A primer is limited to apply on the differentiation of only crude herbal plants. To construct more sensitive and unique primers on the genus Panax, ginseng-specific DNA profile (350 bp) that was amplified by OP-5A primer were inserted in a plasmid vector in the TA cloning method and sequenced. We designed the PCR primers (Forward: 5＂-AGGGGTCTTGCTAT AGCGGAAC-3＂, Reverse: 5＂-AGTCTTAATTTCATATTTTCGTATG-3＂) and identified the unique ginseng band (350 bp) in commercial granule products including ginseng extracts as well as crude ginseng plants by nascent PCR.(omitted)

• Korean journal of applied entomology
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• v.47 no.1
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• pp.37-44
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• 2008

Oriental fruit moth, Grapholita molesta, is a serious pest on apples. To control this pest in an environmentally friendly method, mating disruption strategy using sex pheromone has been developed. Area-wide application of mating disruption has been needed to be effective, with little understanding on how much size of apple cultivating area should be treated in one time application of the mating disruption technique. On this matter, we needed to determine a minimal mating active zone of G. molesta that should be applied with mating disrupters to be effective. Molecular markers to discriminate a specific population should be developed to trace population migration for reproductive behaviors. Here we developed two effective molecular markers using random amplified polymorphic DNA (RAPD) technique. Different field populations of G. molesta, based on locations and seasons, were analyzed with these markers. In a specific location, G. molesta populations varied in genetic composition with different seasons. Different local populations showed differential variation according to their relative distances among apple orchards. In overall, genetic variation among different populations became lessen with progression of seasons.

• The Korean Journal of Mycology
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• v.25 no.3 s.82
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• pp.202-208
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• 1997

In this study, we evaluated the use of RAPD method to discriminate among strains of Fusarium species including F. oxysporum and f. sp. of F. oxysporum. As a result of the amplication, fifteen primers showed total 180 bands ranging from 0.2 to 3 Kb. Among those 180 bands, 126 polymorphic bands were used for bionominal matrix code (0, 1), and UPGMA dendrogram analysis. Fusarium oxysporum isolate 355 showed high similarity with F. oxysporum isolate 358 at 0.9603. Fusarium roseum isolate 87 and F. oxysporum isolate 358, F. o. f. sp. lycopersici isolate 69 and F. o. f. sp. melongena 68 showed low similarity of 0.3809. Fusarium oxysporum isolate 361 and F. o. f. sp. raphani isolate 218 showed similarity of 0.8730, F. oxysoprum isolate 354 and unidentified Fusarium sp. isolate 228 showed similarity matrix of 0.7936, and F. roseum isolate 87 and F. o. f. sp. raphani isolate 57 showed similarity matrix of 0.5873.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.30 no.3
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• pp.388-392
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• 1997

The random amplified polymorphic DNAs (RAPD) technique was used to characterize seven isolates of the green seaweed Ulvales collected from Songjeng, Haeundae, Jumunjin, Dadaepo and Wando in Korea. Total DNA was extracted by the LiCl extraction method from thalli of green seaweed. The extracted DNA (3 ng) in $25{\mu}\ell$ reaction volume was amplified by 45 cycles of the polymerase chain reaction with arbitrary primers. Thirty-four primers resulted in 1227 PCR products ranged 240 bp to 1.5 kb of both conserved and polymorphic bands. Genetic similarities of the seven isolates calculated by Jaccard's equation were ranged from $7\%\;to\;36\%$. Monostroma nitidum (Wando) was shown to be most distantly related with the others based on genetic similarity and did not produce the amplified band of 630 bp, common in Ulvales using primer OPB-01 (CATCCCCTG).

• The Korean Journal of Mycology
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• v.38 no.1
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• pp.34-39
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• 2010

We have attempted to verify 30 strains of Hypsizigus marmoreus from various mushroom stocks in Korea using random amplified polymorphic DNA (RAPD) methodology. Chromosomal DNAs of them were extracted and subjected to PCR analyses with 3 random primers. Each PCR produced approximately 30 distinct PCR bands with the size from 200 bp to 3000 bp. A dendrogram was acquired using the unweighted pair-group method with arithmetic average (UPGMA) clustering methodology on the basis of the DNA band pattern. The analysis revealed that 30 strains of H. marmoreus were clustered into two distinct clusters. Cluster 1 contained 3 subgroups while the cluster 2 consisted of rather diverse strains. Interestingly, Hm3-10, a wild strain collected from Deog-Yu mountain, was not included in either clusters, indicative of uniqueness of this strain. We nextly attempted to develop strain-specific DNA markers to verify a specific strain. A unique band in the RAPD gel lane of Hm0-4 was extracted and its sequence was determined. PCR with a primer set from the determined sequence revealed that the primer set gave a 250 bp DNA band only for Hm0-4, indicating that this approach works well for the strain-specific identification of H. marmoreus.

• FLOWER RESEARCH JOURNAL
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• v.16 no.1
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• pp.28-35
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• 2008

Eleven accessions of Hedera helix, three accessions of Hedera rhombea, one accession of Fatshedera lizei, and one accession of Fatsia japonica were collected and their genetic diversity was measured by using 10 RAPD primers. Approximately ninety seven percentage of polymorphism was detected, because broad germplasm, three genus, was used. Total 97 bands were scored and a dendrogram was constructed by using an UPGMA method. Accessions belonging to Hedera helix tightly clustered in one group: eight accessions showed extremely narrow genetic differences and the other three accessions also showed significant similarity. Despite of their genetic similarity, they showed morphological variations. The morphological variation with a narrow genetic differences indicated that the ivy cultivars have been indeed developed from a mutation breeding program. Genetically most unrelated Fatsia japonica showed a genetic distance of 0.63 on the average between other species. An accession from Fatshedera lizei developed by crossing between two genus, Hedera helix and Fatsia japonica, was allocated together with accessions from Hedera rhombea.

• Korean Journal of Breeding Science
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• v.42 no.5
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• pp.525-533
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• 2010

In this study, random amplified polymorphic DNA (RAPD) and simple sequence repeat (SSR) analyses were utilized for evaluation of genetic diversity of 34 Korean bred and introduced apple cultivars. Thirty-seven RAPD primers detected a total of 193 polymorphic bands (36.2%) with an average of 5.6. Twenty-six SSR markers generated a total of 112 alleles with an average 4.3 alleles per locus. Genetic diversity of 34 cultivars estimated by polymorphic information content (PIC) value ranged from 0.536 (CH03d12) to 0.952 (CH04c06) with an average of 0.843. By UPGMA (unweighted pair-group method arithmetic average) cluster analysis with 305 polymorphic bands, the apple cultivars were classified four groups by similarity index of 0.640. The 'Seokwang' was included in group I. Group II consisted of 12 cultivars which have 'Golden Delicious' in their pedigree, with the exception of 'Spur Earliblaze' and 'Jonathan'. Group III included 13 cultivars which have usually 'Fuji' in their ancestry and bud sport of 'Fuji' cultivars. Group IV consisted of 8 cultivars with 'Hongro', 'Gamhong', and 'Saenara'. Similarity values among the tested apple cultivars ranged from 0.529 to 0.987, and the average similarity value was 0.647. The similarity index was the highest (0.987) between 'Hwarang' and 'Danhong', and the lowest (0.529) between 'Seokwang' and 'Hwarang'. The genetic relationships among the 34 studied apple cultivars were basically consistent with the known pedigree.

• The Korean Journal of Mycology
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• v.26 no.3 s.86
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• pp.365-372
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• 1998

In this study, we evaluated the genetic relationships of fourty seven Pestalotiopsis theae isolates collected from diseased sweet persimon in various places in southern part of Korea using RAPD (Randomly Amplified Polymorphic DNAs) method. As a result of the amplification, eight primers showed total of 86 bands ranging from 0.3 Kb to 3.2 Kb. Among those 86 bands, 84 polymorphic bands were used for bionominal matrix code (0, 1), and UPGMA dendrogram analysis. Similarities among the compared isolates ranged from below 60% to more than 95%. Most of the compared isolates showed $50{\sim}80%$ similarities. The number of isolate pairs which showed more than 80% similarity were 248. The number of isolate pairs which showed $50{\sim}80%$ similarity were 789, and the number of isolate pairs which showed below 50% similarity were 21. Isolate SP-21 (No.9) showed below 50% similarity with all the isolates compared. At 50% similarity level, all the isolates compared, except isolate SP-21 (No.9), were included in one big group. At 65% similarity level, all the isolates compared, except isolate SP-21 (No.9), were divided into three different groups. At 75% similarity level, all the isolates compared, except isolates SP-47 (No. 23) and SP-21 (No.9), were divided into six different groups.

• Journal of Microbiology
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• v.44 no.1
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• pp.106-112
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• 2006

In this study, the genetic diversities of multi-resistant Salmonella typhimurium (ST) isolates were analyzed via the application of both pulsed field gel electrophoresis (PFGE) and Polymerase chain reaction (PCR) analysis methods, using 6 kinds of primers (REP, ERIC, SERE, BOX, P-1254 and OPB-17). And their discriminative abilities (DA) were also compared in order to determine the most effective and reliable analysis method. 118 S. typhimurium isolates, cultured from diverse animals and human patients in Korea beginning in 1993, were analyzed and subjected to a comparison of Simpson's index of diversity (SID), using both PFGE and PCR methods. PFGE by XbaI enzyme digestion allowed for discrimination into 9 pulsotypes, with high SID values (0.991) on the genomic DNA level. This shows that PFGE is a very discriminative genotypic tool, and also that multiple clones of S. typhimurium isolates had existed in domestic animals and humans in Korea since 1993. However, we could ultimately not to trace the definitive sources or animal reservoirs of specific S. typhimurium isolates examined in this study. Depending on the SID values, the combined method (7 kinds of method) was found to be the most discriminative method, followed by (in order) SERE-PCR, REP-PCR, ERIC-PCR, PFGE & OPB-17 (RAPD), P-1254 (RAPD), and BOX-PCR at the $80\%$ clone cut-off value. This finding suggests that the REP-PCR method (which utilizes 4 primer types) may be an alternative tool to PFGE for the genotyping of S. typhimurium isolates, with comparable cost, time, and labor requirement. The establishment of a highly reliable and discriminatory method for epidemiologic analysis is considered necessary in order for researchers to trace the sources of specific pathogens and, consequently, to control and prevent the spread of epidemic S. typhimurium isolates to humans.

• Asian-Australasian Journal of Animal Sciences
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• v.19 no.11
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• pp.1519-1523
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• 2006

Through a screening and selection approach method, decamer random markers were used in a technique called random amplified polymorphic DNA (RAPD) assay with 252 genomic DNAs isolated from four major Haimen chicken populations: Rugao (62), Jiangchun (62), Wan-Nan (63) and Cshiqishi (65). A total of 3-score decamer random primers (S241-S260, S1081-S1100 and S1341-S1360) were employed in the preliminary RAPD-polymerase chain reaction (RAPD-PCR) assay with 50 random template DNA samples from all the populations. Four (6.67%) of the primers that produced obvious polymorphic patterns, interpretable and reproducible bands were selected and used with both the individual DNAs from each population and with pooled DNA samples of the four populations in subsequent analyses. The selected primers produced a total of 131 fragments with molecular size ranging from 835 to 4,972 base pairs (bp) when used with the individual DNAs; 105 (80.15%) of these fragments were polymorphic. With the pooled DNAs, 47 stable and characteristic bands with molecular size ranging from 840 to 4,983 bp, of which 23 (48.94%) polymorphic, were also generated. The band-sharing coefficient (BSC) calculated for the individuals in the population and among populations of bulked samples was between 0.8247 (Rugao) and 0.9500 (Cshiqishi); for pairwise populations, it was between 0.7273 (Rugao vs. Wan-Nan) and 0.9367 (Jiangchun vs. Cshiqishi) chicken populations. Using the BSC for individual and pairwise populations, the Nei's standard genetic distances between the chicken populations were determined and ranged from 0.0043 (Jiangchun vs. Cshiqishi) to 0.1375 (Rugao vs. Cshiqishi). The reconstructed dendrogram linked the Jiangchun and Cshiqishi chickens as closely related populations, followed by Wan-Nan, while the Rugao was the most genetically distant among the populations.