• Journal of Life Science
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• v.24 no.4
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• pp.428-436
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• 2014

Loop-mediated isothermal amplification (LAMP) technique relies on autocycling strand displacement DNA sysnthesis without template denaturation steps under isothermal conditions. LAMP has more advantages than modern PCR, as it requires only basic laboratory equipment like an isothermal water bath, oven, and heating cabinet. Hence, in this study, five random primers were designed with Streptococcus parauberis, shikimate kinase Arok gene sequences (Genbank accession number: CP0024711). Two primers were selected based on the ladder pattern. Other optimum reaction conditions like temperature, time, and sensitivity were established and confirmed with conventional SYBR-green PCR. Results confirmed that the designed random primers were species specific, without any non-target DNA amplification under isothermal conditions. Hence, this study suggests that LAMP techniques could be used in the diagnosis of fish pathogen, specifically S. parauberis.

• Journal of Life Science
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• v.21 no.10
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• pp.1434-1442
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• 2011

A genetic variation within 29 strains of F. velutipes was analyzed by internal transcribed spacer (ITS) sequence analysis and random amplified polymorphic DNA (RAPD). Seven hundred and twenty base pairs were sequenced during the analysis of the ITS region, but no significant variation was observed among the 29 strains of F. velutipes. Sixteen out of 40 random primers amplified polymorphic RAPD fragment patterns. The polymorphic levels of RAPD bands by some primers (OPA-2,4,3,9,10,20) were very high in all 29 strains, with 3,030 fragments ranging between 200 and 2,000 bp. Intraspecific genetic dissimilarity of the 29 strains was calculated to range from 3.3% to 45% by Nei-Li's method using these 3,030 RAPD bands. The genetic variation among Korean strains was relatively high, with dissimilarities ranging between 17% and 38.6%. In the Neighbor-Joining analysis using the genetic dissimilarities based on RAPD, all 29 strains were classified into 5 clusters. Strains in each cluster showed specific characteristics according to their origin and strains. These results suggested that OPA and OPB primers could be used for developing molecular genetic markers and screening of unidentified (F. velutipes) strains.

• Journal of Sericultural and Entomological Science
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• v.38 no.1
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• pp.7-12
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• 1996

Reproducible the random amplified polymorphic DNAs(RAPDs) patterns were obtained in the two silkworm strains(J111, Galwon) by adjusting concentration optimized of Taq DNA polymerase(one unit), dNTP(200$\mu$M), MgCl2(1.5mM) and template DNA(30ng). In addition, anealing temperature ranging 35$^{\circ}C$ to 42$^{\circ}C$ by the adjusted condition was investigated and fixed at 35$^{\circ}C$ in this study. Variation among individuals and between male and female of Jam 113 strain was not authorized. DNA polymorhpisms among silkworms were authorized by five RAPD markers using OPM04 random primer. Using the primer showing polymorhpims between parents(J111, Galwon) in thirty three individuals, RAPD-PCR for F2 analysis was performed and segregated 3 : 1 in the F2 population. Consequently, RAPDs detected in the parents were obtained as genetic markers, which can be used for construction of genetic map for this industrially particular insect, silkworm Bombyx mori

• Korean Journal of Medicinal Crop Science
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• v.12 no.1
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• pp.31-35
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• 2004

The genetic variation and intraspecific relationships between 10 individuals of seven cultivars and one Ulleungdo native of Wasabia japonica were investigated using RAPD (Randomly Amplified Polymorphic DNA) analysis. The 21 primers out of 50 random primers were amplified for all tested plants. The 68 (47.2%) among 144 bands derived from 21 primers showed polymorphism, and 3.2 bands per primer were observed. Number of bands per primer was ranged from 2 to 13, and average numbers were 6.8. The phenograms for 11 analyzed individuals by RAPD markers were not matched well with those of the result by morphological characters since they were clustered monophyletic at the similarity coefficient value ranged from 0.81 to 0.96. The Ulleungdo native individual was clustered sister to Daruma, Simanesairal, Sawa, and Hujidaruma cultivars. The RAPD markers were not useful to evaluate the intraspecific variations in Wasabia japonica cultivars, therefore need to more specific molecular phylogenetic characters such as AFLP technology and gene sequence of nuclear and chloroplast DNA.

• Korean Journal of Microbiology
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• v.35 no.2
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• pp.107-114
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• 1999

To assess genetic diversity amoug 21 strains from sixleen Frrsn~i~nn species , we used RAPD(rando1n amplified pol.ymorphic DNA) analysis based on PCR(po1ymerase chain reaction). Eleven primers showing Ule polymorphism were chosen from the 40 random pnmers-tcstcd. A total of 263 polymorphic bands were generated by the primers and the size of amplified DNA fragments ranged from 0.1 lo 3.0 kb. Sirnilku-it), coefficients between strains were calcnlatcd, and UPGMA cluster analysis was used to generate a dendrogram showing relationships among them. The results from RAPD-PCR analysis were grouped into four main groups at the si~nilarity level of 0.627.

• The Korean Journal of Food And Nutrition
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• v.17 no.3
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• pp.254-259
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• 2004

Rapid detection of foodbome pathogens is becoming increasingly important. The requirement for faster, more reliable tests has lead to the development of a wide range of rapid methods. Among these methods, the use of systems based on nucleic acid based detection has been increasing since they offer advantages of reduction in test time and more reliable detection or identification. Random Amplification Polymorphic DNA(RAPD) method has been used to fingerprint foodbome microorganisms; Listeria monocytogenes. In this study, 10-mer primer OPG-13(5'-CTCTCCGCCA-3') was used to generate RAPD-PCR for detection of pathogenic L. monocytogenes of Listeria spp. Among 20 primers tested, OPG-13 showed on acceptable result for the differentiation of a pathogenic Listeria from non-pathogenic microorganisms. Pathogenic Listeria, L. monocytogenes(ATCC 15313, 19111, 19112, 19113) showed two bands for 700 bp and 1,500 bp while non-pathogenic bacteria, L. ivanovii, L. grayi, L. murrayi, L. innocua, L. welshimeri, and L. seeligeri had only one band sizing from 2,000 to 2,300 bp. This RAPD method proved to be a valuable to gain important information on sources of pathogenic bacteria in food industry.

• The Korean Journal of Mycology
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• v.25 no.3 s.82
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• pp.176-190
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• 1997

Sixty-three isolates of Lentinus edodes obtained from Korea were used to assess the genetic similarity by isozyme polymorphisms and random amplified polymorphic DNA patterns. The activities of esterase, peroxidase and acid phosphatase displayed 10, 7 and 3 distinct isozyme patterns, respectively. By combining the isozyme patterns obtained with the 3 enzymes, every isolate showed its own distinct electrophoretic phenotypes. A distance matrix calculated between all pairs of 63 electrophoretic phenotypes based on the presence or abscence of isozyme bands were analyzed by the group-average method. Results of the cluster analysis assinged the 63 phenotypes into six major groups. In the analysis of random amplified polymorphic DNA patterns, all isolates of Lentinus edodes were devided into five RAPD groups.

• Molecules and Cells
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• v.26 no.3
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• pp.250-257
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• 2008

Microsatellites or simple sequence repeats (SSR) are widely distributed in eukaryotic genomes and are informative genetic markers. Despite many advantages of SSR markers such as a high degree of allelic polymorphisms, co-dominant inheritance, multi-allelism, and genome-wide coverage in various plant species, they also have shortcomings such as low polymorphic rates between genetically close lines, especially in Capsicum annuum. We developed an alternative technique to SSR by normalizing and alternating anchored primers in random amplified microsatellite polymorphisms (RAMP). This technique, designated reverse random amplified microsatellite polymorphism (rRAMP), allows the detection of nucleotide variation in the 3' region flanking an SSR using normalized anchored and random primer combinations. The reproducibility and frequency of polymorphic loci in rRAMP was vigorously enhanced by translocation of the 5' anchor of repeat sequences to the 3' end position and selective use of moderate arbitrary primers. In our study, the PCR banding pattern of rRAMP was highly dependent on the frequency of repeat motifs and primer combinations with random primers. Linkage analysis showed that rRAMP markers were well scattered on an intra-specific pepper map. Based on these results, we suggest that this technique is useful for studying genetic diversity, molecular fingerprinting, and rapidly constructing molecular maps for diverse plant species.

• Biomedical Science Letters
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• v.24 no.3
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• pp.230-238
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• 2018

Human enteric Adenovirus-41 (HuEAdV-41) causes gastroenteritis, which detected by the polymerase chain reaction (PCR) base diagnostic system for clinical, food, environmental, fish and shellfish samples. We developed improved PCR and nested PCR primer set which had high specificity, sensitivity and reduced times. In this study, we compared seventeen conditions reported in the previous study that was using the PCR based HuEAdV-41 detection system, and non-enteric Adenovirus were detected in nine conditions. The most sensitive detection condition was up to 25 copies however it took 184 minutes of PCR reaction time. In this study, the PCR primer set developed had same level of sensitivity, it reduced the time of detection for clinical, food and seafood samples to 112 minutes. Developed nested PCR primer set needed 112 minutes but detected up to approximately 1 copy. In addition, developed PCR and nested PCR primer set was validated with twenty samples of underground water at random, of which ten samples showed specific band without non-specific reaction. We expect this study will be used to diagnose HuEAdV-41 from various samples.

• Korean Journal of Agricultural Science
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• v.26 no.2
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• pp.39-48
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• 1999

This study was carried out to investigate the difference and genetic similarity at the level of molecular genetics. Genomic DNA was extracted from blood of Holstein, Korean cattle, Charolais, and hybrid between Korean cattle and charolais and RAPD(random amplified polymorphic DNAs) was analyzed by PCR(polymerase chain reaction). After genetic similarity value from different breeds are analyzed, genetic similarity was estimated by UPGMA(unweighted pair-group method using average). The results obtained from this study can be summarized as follows: 1. When genomic DNA which was extracted from different breeds was subjected to electrophoresis on 1.5% agarose gel, bigger than 12.2kb was appeared. Ratio by absorbance of $A_{260}/A_{280}$ was 1.75~2.10, indicating that genomic DNA was quite pure for RAPD analysis. 2. Different band patterns by RAPD were appeared according to the breeds in cattle. The best primer used to distinguish Holstein from other breeds was 5'-GAC CGC TTG T-3'. 3. A 340bp fragment was amplified in $33.0^{\circ}C$ of annealing temperature for the Holstein and Charolais breeds, but any amplification was not occurred in this annealing temperature for Korean cattle and hybrid. In addition, a 340bp fragment was amplified in $37.5^{\circ}C$ of annealing temperature for the Holstein and Korean cattle, but any amplification was not occurred in this annealing temperature for Charolais and hybrid. For the reaction of PCR. $37.5^{\circ}C$ and $33.0^{\circ}C$ of annealing temperature was shown to be best for genetic marker identification from Holstein, Charolais, and hybrid between Korean cattle and Charolais. 4. When genetic similarity from different breeds are analyzed at the both temperature of $33.0^{\circ}C$ and $37.5^{\circ}C$, the genetic similarity value of Holstein and Korean cattle, Holstein and Charolais, Korean cattle and Charolais, and Korean cattle and hybrid were 0.666~0.777, 0.615~0.666, 0.400~0.461 and 0.857~0.888, respectively. 5. It could be concluded that different breeds are capable of distinguishing by RAPD used random primer 5'-GAC CGC TTG T-3', genetic similarity from different breeds was appeared the higher genetic similarity value of Korean cattle and Charolais than that of Holstein between Korean cattle and Charolais by UPGMA.