• Journal of Biosystems Engineering
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• v.28 no.5
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• pp.429-438
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• 2003

This study exploits the robot system which is necessary in gene study and bio-technology industry. As well, a DNA chip, which of use has been increased recently, can be manufactured with this system. The robot consists of a device spotting DNA on the silylated slide, a well plate, a bed for fixing well plates, devices of washing and drying the pin in DNA spotting .device, a distillation-water vessel, and a discharge vessel of wash water. We made the period of sticking DNA to the pin on the well plate to be 15 seconds. The spot size of DNA was set to be 0.28 mm on the average by bringing the slide into contact with pin during 1 second. If DNA is spotted in minimum space possible about 0.32mm, this system can stick about 8,100 DNA spots on the well plate with this rate. Analyzing the procedure: Movement starts, Pin washes, dries, and smears DNA on the well plate. Spotting DNA onto 12 chips took 2 minutes and 50 seconds.

• Korean Journal of Agricultural Science
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• v.30 no.1
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• pp.76-88
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• 2003

This study exploits the robot system which is necessary in gene study, bio-technology industry. As well, it can achieve the job of DNA chip manufacturing whose use rate has been increased recently. The robot consists of DNA spotting device for spotting DNA on the silylated slide and well plate, bed for fixing well-plate, washing & drying device of washing and drying the pin part of DNA spotting device, distillation-water vessel, and discharge vessel of wash water. We made the term of sticking DNA to the pin on well plate to be 15 seconds. The spot size of DNA was set to be 0.28 mm on the average by bringing the slide into contact with pin for 1 second. At this rate, if DNA is spotted in the minimum space possible of about 0.32mm, it can stick about 8,100 DNA spots on the well plate. Analyzing the procedure: Movement starts. Pin washes, dries, and smears DNA on the well plate. Spots DNA onto 12 chips takes 2 minutes and 50 seconds.

• Proceedings of the Korean Society of Fisheries Technology Conference
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• 2003.05a
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• pp.311-312
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• 2003

Microarray를 해석하는데 있어서 방해가 되는 요인에는 여러 가지가 있을 수 있다. 예를 들면 probe DNA의 농도, spotting 시의 습도, spot의 크기 및 모양, slide blocking 과정, lab리ing technique, hybri야zation 온도와 시간 그리고 background signal 등이 그것이다(Hegde et at., 2000). 본 연구에서는 전보에서 선정된 rhabdovirus의 96개 probe로 실제 oligonucleotide chip을 만들어 spot의 모양이나 크기에 관계되는 spotting 조건을 확립하고, slide blocking과 slide washing에 따른 background signal을 낮추기 위한 과정을 도입하였다. (중략)

• Proceedings of the Korean Society of Applied Pharmacology
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• 1998.11a
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• pp.183-183
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• 1998

Telomerase synthesizes telomeric DNA repeats onto chromosome ends de novo. Telomerase activation and telomere shortening in human somatic cells have been implicated in cell tumorigenesis and immortalization. In order to find the potential inhibitors against telomerase activitiy which can be used as potential anticancer agents, we screened about 100 kinds of natural products after partition into n-hexane, ethyl acetate and aqueous layers from methanol extracts. The inhibitory effects of these materials against telomerase enzyme activity were tested in 293T cell culture using telomeric repeat amplification protocol(TRAP). The incorporation of $\^$32/P-dGTP into amplified DNA was measured by adsorption to Whatman DE81 paper instead of using TRAP assay for screening the extracts of natural products. Strong effective compounds were not found in this study but DE81 filter spotting method may be a useful model for the screening. Some of the compounds which showed somewhat inhibitory effects had cytotoxic effects also.

• Proceedings of the Korean Society for Agricultural Machinery Conference
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• v.10 no.2
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• pp.337-342
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• 2005

• The Journal of the Korea institute of electronic communication sciences
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• v.8 no.12
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• pp.1875-1882
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• 2013

Micro-array technology contributes numerous achievements such as ordering of gene network and integration of genomic. This technology is well established as means for investigating patterns of gene expression. DNA micro-arrays utilize Affymetric chips where a large quantity of DNA sequences may be synthesized. There are two general type of conventional DNA array spotter: contact and piezoelectric. The contact technology used spotting pin technology to make contact with the glass slide surface. This may caused damage or scratches to the surface matrix where protein will be contaminated and may not bind specifically. Piezoelectric technology available at this present time on the other hand requires the analyzer to print the result that can only be done within the laboratory despite of mass production. Therefore, in this paper, high-throughput technology is developed for providing greater consistency in feature spot without touching the glass slide surface.

• BMB Reports
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• v.37 no.5
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• pp.546-551
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• 2004

The increasing pace of development in molecular biology during the last decade has had a direct effect on mass testing and diagnostic applications, including blood screening. We report the model Microarray that has been developed for Hepatitis B virus (HBV) and Hepatitis D virus (HDV) detection. The specific primer pairs of PCR were designed using the Primer Premier 5.00 program according to the conserved regions of HBV and HDV. PCR fragments were purified and cloned into pMD18-T vectors. The recombinant plasmids were extracted from positive clones and the target gene fragments were sequenced. The DNA microarray was prepared by robotically spotting PCR products onto the surface of glass slides. Sequences were aligned, and the results obtained showed that the products of PCR amplification were the required specific gene fragments of HBV, and HDV. Samples were labeled by Restriction Display PCR (RD-PCR). Gene chip hybridizing signals showed that the specificity and sensitivity required for HBV and HDV detection were satisfied. Using PCR amplified products to construct gene chips for the simultaneous clinical diagnosis of HBV and HDV resulted in a quick, simple, and effective method. We conclude that the DNA microarray assay system might be useful as a diagnostic technique in the clinical laboratory. Further applications of RD-PCR for the sample labeling could speed up microarray multi-virus detection.

• Journal of Animal Science and Technology
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• v.44 no.6
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• pp.653-660
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• 2002

We considered KIT gene as a candidate gene for the white-spotting pattern in cattle. This study was carried out to detect genetic variation of c-KIT receptor gene and to investigate association between the mutation and the white-spotting pattern in cattle. PCR-RFLP analysis within intron 6 of c-KIT receptor gene were performed with 8 cattle breeds including Hanwoo, Angus, Brown Swiss, Charolais, Hereford, Holstein, Limousin and Simmental. When PCR product of approximately 2,440 bp including intron 6 of c-KIT receptor gene was sequenced, four nucleotide substitutions were found within intron 6 of the bovine c-KIT receptor gene. In PCR-RFLP analysis, three alleles (A, B and C), two alleles (A and B) and two alleles (A and B) at each locus were identified by MspⅠ, BsrBⅠ and NdeⅠ, respectively. Although frequencies of allele at each locus were different among cattle breeds, we could not get any evidence related with white or white spotting phenotypes in these mutations on intron 6 of c-KIT receptor gene. However, we can not entirely exclude the possibility that c-KIT receptor gene is responsible for white spotting phenotype in cattle. Thus, further studies need to detect other mutations in c-KIT receptor gene and to test association of those mutations and coat color phenotypes in cattle.

• Biotechnology and Bioprocess Engineering:BBE
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• v.10 no.5
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• pp.432-443
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• 2005

Microarray technology has contributed Significantly to the understanding of bacterial genetics and transcriptional regulation. One neglected aspect of this technology has been optimization of microarray-generated signals and quality of generated information. Full genome microarrays were developed for Clostridium acetobutylicum through spotting of PCR products that were designed with minimal homology with all other genes within the genome. Using statistical analyses it is demonstrated that Signal quality is significantly improved by increasing the hybridization volume. possibly increasing the effective number of transcripts available to bind to a given spot, while changes in labeled probe amounts were found to be less sensitive to improving signal quality. In addition to Q-RT-PCR, array validation was tested by examining the transcriptional program of a mutant (M5) strain lacking the pSOL1 178-gene megaplasmid relative to the wildtype (WT) strain. Under optimal conditions, it is demonstrated that the fraction of false positive genes is 1% when considering differentially expressed genes and 7% when considering all genes with signal above background. To enhance genomic-scale understanding of organismal physiology, using data from these microarrays we estimated that $40{\sim}55%$ of the C. acetobutylicum genome is expressed at any time during batch culture, similar to estimates made for Bacillus subtilis.

• The Plant Pathology Journal
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• v.19 no.4
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• pp.210-216
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• 2003

This study examined the existence of genetically diverse population of Cucumber mosaic virus (CMV), known as quasispecies, from lily, Nicotiana benthamiana and from purified virions. Based on the conserved sequences of CMV lily isolates in intergenic region (IR) on RNA3, the genetic variation of IR from three different sources was investigated by a specific restriction endonuclease hydrolysis of amplified reverse transcription-polymerase chain reaction (RT-PCR) products using virus-specific primers, and was compared with IR sequences. The IR nucleotide sequences of CMV lily isolates were highly conserved, however, quasispecies was detected from all three sources in low level, containing sub-populations of RNA3. These subpopulations of RNA3 were inoculated onto zucchini squash by in vitro transcripts from corresponding full-length cDNA clones together with Eny RNA1 and 2 transcripts. The systemic symptom of zucchini plants infected by these quasispecies was chlorotic spotting, which was milder than severe mosaic and stunt symptom caused by Eny-CMV. The severity of symptom was correlated with RNA accumulation of viruses. These results suggest that the genome of CMV lily isolates consists of quasispecies populations.