• Korean Journal of Medicinal Crop Science
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• v.8 no.3
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• pp.266-273
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• 2000

The optimal conditions of PCR components for the random amplification of genomic DNA were $20\;ng/20{\mu}l$ in template DNAs, 250 mM in dNTP, 10 pM in primer $1.0unit/20{\mu}l$ in Taq DNA polymerase respectively with the annealing temperature at $36^{\circ}C$, respectively. Twelve local lines were divided into 3 groups by the coefficients of 107 polymophic bands by Jaccard and Nei. The coefficients value of group I including Chongup # 1, Seochon # 1, Andong # 1, Chinan # 1, and Danyang # 1 ranged from 0.27 to 0.05 and those of group II including Suwon # 2, Chunchon # 1, Japan # 3, Danyang#2 and $F_1$ (Variety Jihwang $1{\times}$ Seohchon) ranged from 0.29 to 0.11. While, Jihwang 1 originated from China and Japan # 1 in group III showed a distant genetic relationship to Korean local lines.

• Horticultural Science & Technology
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• v.19 no.1
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• pp.29-33
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• 2001

This study was carried out to discriminate potato cultivars and breeding lines by specific molecular markers using random amplified polymorphic DNA (RAPD) analysis. The genotypes of potatoes used for analysis were eight cultivars and five breeding lines. Some of those show much phenotypic resemblances among them because 'Jopung', 'Daekwan70', 'Gawon', and 'Daekwan72' have immediate parental relationship with 'Superior', 'Irish Cobbler', 'Namsuh', and 'Atlantic', respectively. So, there are many difficulties to distinguish the varieties by the morphological characteristics. Three URP primers, URP2, URP4, and URP8 were selected for promising primers to discriminate potato genotypes or cultivars. The three URP primers were shown very high reproducibility because of the relatively high annealing temperature and long primer size. Although the results of similarity analyses did not always reflect the genetic relationship between potato varieties, the reproducible pattern of amplified DNA bands by URP primers showed possibility for molecular markers for discrimination of potato genotype or cultivar.

• Fisheries and Aquatic Sciences
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• v.6 no.1
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• pp.13-19
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• 2003

Genomic DNA from the muscle of marsh clam (Corbicula leana) from Gochang, Muan and a Chinese site was extracted to identify genetic differences and similarity by randomly amplified polymorphic DNAs-polymerase chain reaction (RAPD- PCR). Out of 20 primers, seven primers produced amplified fragments which were consistently polymorphic. A total of 1,246 amplified products were produced of which 530 were polymorphic $(42.5\%)$. The number of polymorphic bands produced per primer varied from 40 to 122 with an average of 75.7 in marsh clam from Gochang. 3.28 of the 23.0 polymorphic bands per lane were found to be polymorphic. Also, about $4.34\%$ of total polymorphic bands were specific to marsh clam from Gochang. The major common bands of 0.28 kb generated by primer OPB-15 (GGAGGGTGTT) were present in every individuals, which were polymorphic. This common bands in every individuals should be diagnostic of specific strains, species and/or their relatedness. Primer OPB-19 (ACCCCCGAAG) produced the highest number of 12 specific bands. The intra-population variation was revealed in the band patterns identified by this primer. The random primer OPB-12 (CCTTGACGCA) yielded the amplified fragments which were consistently polymorphic between the marsh clams from Gochang and from Muan. This primer produced a total of 77 polymorphic bands: 31 bands from Gochang, 14 from Muan and 32 from the Chinese populations. An average of polymorphic bands were 1.8 from Gochang and 2.5 from the Chinese populations. This value obtained from the Chinese population was higher than those from the two domestic populations. Generally, the RAPD polymorphism generated by these primers may be useful as a genetic marker for strain or population identification of marsh clam.

• Journal of Life Science
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• v.14 no.1
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• pp.110-114
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• 2004

The optimized RAPD-PCR conditions, that can be utilized as a basic information for analysis of the gelletic characteristics were developed for genetic analysis of four mulberry varieties, named Milsung, Chungil, Suil, and Hansung using a primer, OPY15 (5'-AGTCGCCCTT-3') from Operon company. We tested several different factors for best PCR condition including concentrations of DNA, primer, Mgclu annealing temperature, number of PCR cycle, and prosence/absence of pre-heating time at the begining of PCR reaction in the $25 \mul$volume. The best RAPD profiles were obtained using 50 ng of DNA, 1 $\mu$M of primer, $1 \mum$of $MgCl_2\;,45^{\circ}C$ of annealing temperature and an absence of pre-heating time. An establishment of the stable and reproducible RAPD-PCR conditions are expected to be useful for the subsequent RAPD-related investigation, such as genetic characterization of the mulberry varieties, re-establishment of phylogenetic relationships and development of new varieties.

• 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 Mycology
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• v.26 no.4 s.87
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• pp.574-582
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• 1998

Five basidiocarps of Pholiota species have been collected from the areas of BubJu Temple for last two years, and identified to those of P. adiposa or Pholiota species. The taxonomy of these basidiocarps with the morphological aspects was compared with the analysis obtained from the polymorphisms of PCR-RAPD bands made after reacted with the random primers. The polymorphic variations were observed within the species of the basidiocarps, but not between genomic DNA's of the mycelia obtained and the basidiocarps. Several different bands made from the primers (28 and 36) in PCR-RAPD reactions were identified within the genus of Pholiota and speculated to be specific for the individual basidiocarp of P. adiposa collected. The primers employed here were considered to be very useful for distinguishing the individual isolates or basidiocarps collected from the fields. Also, the basidiospores were obtained from the sporeprints of the above basidiocarps as a simple agar and confirmed with observations of clamp connection under microscopes. The matings of them indicated the 'tetrapolar' type, being different from the 'bipolar' type reported by Japanese basidiocarps of P. adiposa or P. nameko. Based on our work, the edible fungi collected were speculated to be a new breeding resource for those of Pholiota commercialized in Japan.

• 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.

• Korean Journal of Poultry Science
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• v.20 no.4
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• pp.177-188
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• 1993

This study was performed to find out the reasonable sexing methods In the chicken, obtain the basic information for the mechanisms related to chicken sexual differentiation and identify the genes which known to involved in chicken sex differentiation. The chromosome analysis of chicken embryonic fibroblast was a simple method to determine sex of chicken by means of Z and W chromosome identification. The bands of female chicken genomic DNA digested with Xho Ⅰ and Eco RI restriction endonuclease showed to be useful in direct sex determination and these repetitive sequences of Xho Ⅰ and Eco RI families were proposed to be very homologous in their sequences by colony hybridization analysis. Seven of 150 random primers were selected to amplify the W chromosome-specific band by using arbitrary primed PCR and three of them were useful to identify the sex of chicken. To identify the sex differentiation genes in the chicken, PCR for the amplification of ZFY and SRY sequences was performed. ZFY and SRY sequences were amplified successfully in the chicken genome, implying that chicken genome might have the sex-related conserved sequences similar to mammalian ones. The PCR products of ZFY amplification were the same in both sexes, suggesting that these sequences may be located on autosome or Z chromosome. The profile of PCR amplification for SRY sequences showed variation between sexes, but this result was not enough to specify whether the SRY gene in chicken is on the autosome or sex chromosome.

• Journal of Life Science
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• v.16 no.2 s.75
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• pp.345-351
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• 2006

Streptococcus sp. were isolated from cultured flounder (Paralichthys olivaceus) having Streptococcosis during 2004 to 2005 in Jeju Island. Ninety four Streptococcus iniae strains were isolated using biochemical test and multiplex PCR assay. Three genotypes (A, B, C-type) of S. iniae were appeared in the RAPD analysis and they showed international or local genetic polymorphism. Presently, S. iniae having A-type is a dominant S. iniae genotype in Jeju and showed band patterns at about 550, 850, 1000, 1300 and 2000 base pares. In this study, the reported P14 random primer, that used to distinguish serotypes of S. iniae could not be applied to distinguish Jeju island S. iniae's genetic polymorphism.

• Plant Resources
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• v.3 no.3
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• pp.211-218
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• 2000

Intraspecific genetic relationship of 19 variation types of the Yam (Dioscorea alata) classified by their external morphological characteristics such as leaf and tuber shape were assessed by DNA using random and specific primer. Twenty two out of 113 primers (100 random[10-mer] primers, two 15 mer [M13 core sequence, and (GGAT)$_4$ sequence]) had been used in PCR-amplification. Only 12 primers, however, were success in DNA amplification in all of the analyzed plants, resulting in 93 randomly and specifically amplified DNA fragments. The analyzed taxa showed very high polymorphisms(69 bands, 71.0 %), allowing individual taxon to be identified based on DNA fingerprinting. Monomorphic bands among total amplified DNA bands of each primer was low under the 50%. Similarity indices between accessions were computed from PCR(polymerase chain reaction) data, and genetic relationships among intraspecific variations were closely related at the levels ranging from 0.66 to 0.90. These DNA data were not matched well with those of morphological characters since they were divided into two major groups at the similarity coefficient value of 0.70. Therefore, Grouping of species into variation types by mainly morphological charactistics was suggested unreasonable.