• Title/Summary/Keyword: hybridization

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Detection of SNPs using electrical biased method on diamond FETs (다이아몬드 FETs에서 전기적 바이어스 방법을 이용한 단일염기 다형성(SNPs) 검출)

  • Song, Kwang Soup
    • Journal of the Institute of Electronics and Information Engineers
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    • v.52 no.3
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    • pp.190-195
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    • 2015
  • The detection of single nucleotide polymorphisms (SNPs) caused of mutant or genetic diseases is important to diagnosis and medicine. There are many methods have been proposed to detect SNPs. However the detection of SNPs is difficulty, because the difference of energy between complementary DNA (cDMA) and SNPs is very small. In this work, we detect the SNPs using field-effect transistors (FETs) which based on the detection of negative charge of DNA. We bias -0.3 V on the drain-source electrode at the target DNA hybridization process. The efficiency of hybridization and the amplitude of signal decrease by repulsive force between negative charge of DNA and negative bias on the electrode. However, the sensitivity of SNPs increases about 5 times from 1.7 mV to 8.7 mV.

Image Analysis Algorithms for Comparative Genomic Hybridization (분자 세포 유전학 기법에 응용되는 영상 처리 기술)

  • Kim, De-Sok;Yoo, Jin-Sung;Lee, Jin-Woo;Kim, Jong-Won;Moon, Shin-Yong;Choi, Young-Min
    • Proceedings of the KOSOMBE Conference
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    • v.1998 no.11
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    • pp.66-69
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    • 1998
  • Comparative genomic hybridization (CGH) is an important molecular cytogenetics technique that maps abnormal copy number of specific DNA sequence of the chromosome. CGH is based on quantitative digital image analysis of ratio images from fluorescently labeled chromosomes. In this paper, we would like to introduce how recently developed image analysis algorithms are used for CGH techniques. To average the ratio profile of each chromosome, binarization, skeletonization, and stretching of chromosome images have been studied. Developed algorithms have been implemented in the karyotyping system ChIPS commercially developed at Biomedlab Co. Ltd.

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Diagnosis of canine distemper by in situ hybridization (In situ hybridization에 의한 개 디스템퍼의 진단)

  • Cho, Hyeon;Park, Nam-yong;Kim, Yong-hwan;Cho, Kyoung-oh;Park, Hyung-seon;Park, Young-seok;Lee, Bong-joo;Chung, Chi-young;Im, Hyung-ho
    • Korean Journal of Veterinary Research
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    • v.39 no.3
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    • pp.583-592
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    • 1999
  • We have developed the in situ hybridization(ISH) technique for rapid diagnosis of canine distemper(CD) which is the major infectious disease in dogs. In our experiment, we rapidly detected distribution of the specific canine distemper viral genome without disrupting morphology of tissues or cells. Two oligonucleotide probes for ISH were synthesized chemically and labelled 5' end with nonisotopic biotin by DNA synthesizer. The whole procedures of ISH was completed within 1~2 hours using the Microcapillary action system. On histological study, typical cytoplasmic or intranuclear inclusion bodies were observed in the trachea, bronchiole, brain, and urinary bladder with the presence of prominent red positive signals on ISH, indicating specific CDV genome from the paraffin-embedded tissues of infected 13 cases. The results showed ISH can be used as a rapid and effective diagnostic method for diagnosis of CD.

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Histopathological observations and virus detection by in situ hybridization in wild rats intranasally infected with Aujeszky's disease virus isolated in Korea (국내분리 오제스키병 바이러스로 비강접종한 야생집쥐(Rattus norvegicus)의 병리학적 소견 및 in situ hybridization에 의한 바이러스 동정)

  • Song, Geun-seok;Moon, Oun-kyong;Jeong, Chang-geun;Kim, Soon-bok
    • Korean Journal of Veterinary Research
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    • v.39 no.3
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    • pp.531-537
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    • 1999
  • The present study was carried out to investigate the pathogenicity and pathogenesis of wild rats(Rattus norvegicus), trapped in nature, intranasally infected with Aujeszky's disease virus(ADV/NYJ-1-87) by histopathology, immunohistochemistry and in situ hybridization(ISH). Fifteen rats inoculated intranasally were roughened haircoat, anorexia, listlessness, and depression second day after inoculation, and three rats died in 66-72 hours. Eight rats showed severe pruritus at the face that was accompanied by frequent face-washing movements of the forelegs, and then became violent and spasmodic for an hour or until they died. Four rats slowly recovered after showing mild clinical signs of the disease. Microscopic lesions in infected rats were characterized by meningitis, perivascular round cell infiltration, focal gliosis, and neuronal degeneration and necrosis. And intranuclear inclusion bodies were frequently detected in the cerebral cortex and medulla. Positive reaction to ADV by immunohistochemistry and ISH were detected in the following areas : trigermimal ganglion, brain, tonsil, nasal mucosa, spleen, lung and liver. The result has suggested that ADV intranasally infected in wild rats is followed by replication in epithelial calls of nasal mucosa and tonsil, then invade local lymph nodes by way of the lymphatics. It is also believed that the virus invades bipolar olfactory cells and trigerminal ganglion; and then spread into central nervous system.

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High Yield Saponin Production by Mass Cultures of Ginseng Transformed Tissue I. Induction, Culture of Transformed Tissue and Selection of High-Saponin-Producing Clones in Ginseng (인삼 형질전환 조직의 다량배양에 의한 Saponin 고 생산 I. 인삼에서 형질전환 조직의 유도, 배양과 Saponin 고 생산능주 선발)

  • 이정석;고경민
    • KSBB Journal
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    • v.9 no.2
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    • pp.157-164
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    • 1994
  • Hairy root clones of Panax ginseng were established by selection of some hairy roots formed on the leaf, stem and root segments transformed with Agrobacterium rhizogenes strain $A_4$. The transformed roots grew well in MS medium under the dark condition. To confirm the transformation with Ri-T-DNA, dot blot hybridization and opine analysis were Performed. Among four hairy roots induced from different part of ginseng, the HB3 hairy roots were examined for selection of high-saponin-producing clones. Four clones isolated from HB3 hairy root cultures displayed various phenotypes characterized by growth and total saponin content. Maximum growth was obtained for cultures of HB3-10 clone and the content of total saponin was 0.55 wt%. However, higher amount of total saponin was obtained with HB3-2 clone cultures(0.74 wt%) in spite of lower growth. Dot blot hybridization confirmed the introduction of Ri-T-DNA in the plant genome. In the opine test, agropine and mannopine were detected from all hairy root clones.

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Comparison of Hybridization Behavior between Double and Single Strand of Targets and the Application of Asymmetric PCR Targets in cDNA Microarray

  • Wei, Qing;Liu, Sanzhen;Huang, Jianfeng;Mao, Xueying;Chu, Xiaohui;Wang, Yu;Qiu, Minyan;Mao, Yumin;Xie, Yi;Li, Yao
    • BMB Reports
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    • v.37 no.4
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    • pp.439-444
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    • 2004
  • Double stranded targets on the cDNA microarray contain representatives of both the coding and noncoding strands, which will introduce hybridization competition with probes. Here, the effect of double and single strands of targets on the signal intensity and the ratios of Cy5/Cy3 within the same slide were compared. The results show that single stranded targets can increase the hybridization efficiency without changing the Cy5/Cy3 ratio. Based on these results, a new strategy was established by generating cDNA targets with asymmetric PCR, instead of conventional PCR, to increase the sensitivity of the cDNA microarray. Furthermore, the feasibility of this approach was validated. The results indicate that the cDNA microarray system based on asymmetric PCR is more sensitive, with no decrease in the reliability and reproducibility as compared with that based on conventional symmetric PCR.

Hybridization of Quercus aliena Blume and Q. serrata Murray in Korea - Analyses of Morphological variation and Flavonoid chemistry -

  • Park, Jin Hee;Park, Chong-Wook
    • Korean Journal of Environment and Ecology
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    • v.29 no.2
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    • pp.145-161
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    • 2015
  • This research was conducted in order to understand the hybridization between Quercus aliena Blume and Q. serrata Murray in Korea which show wide range of morphological variations within species and interspecific variations of diverse overlapping characteristics caused by hybridization. Morphological analysis (principal components analysis; PCA) of 116 individuals representing two species and their intermediates were performed. As a result, two species were clearly distinguished in terms of morphology, and intermediate morpho-types assumed to be hybrids between the two species were mostly located in the middle of each parent species in the plot of the principal components analysis. There was a clear distinction between two species in trichome distribution pattern which is an important diagnostic character in taxonomy of genus Quercus, whereas intermediate morpho-types showed intermediate state between two species' trichome distributions. Forty-two individuals representing two species and their intermediates were examined for leaf flavonoid constituents. Twenty-three flavonoid compounds were isolated and identified: They were glycosylated derivatives of flavonols, kaempferol, quercetin, isorhamnetin and myricetin. The flavonoid constituents of Q. aliena were five glycosylated derivatives: kaempferol 3-O-galactoside, kaempferol 3-O-glucoside, quercetin 3-O-galactoside, quercetin 3-O-glucoside, and Isorhamnetin 3-O-glucoside. The flavonoid constituents of Q. serrata had 20 diverse flavonol compounds including five flavonoid compounds found in Q. aliena. It was found that there is a clear difference in flavonoid constituents of Q. aliena and Q. serrata. Flavonoid chemistry is very useful in recognizing each species and putative hybrids. The flavonoid constituents of intermediates were a mixture of the two species' constituents and they generally showed similar characteristics to morpho-types. The hybrids between Q. aliena and Q. serrata showed morphologically and chemically diverse characteristics and it is assumed that there are frequent interspecific hybridization and introgression.

The Optimization of Human Sperm Decondensation Procedure for Fluorescence in Situ Hybridization (Fluorescence in Situ Hybridization 시행을 위한 인간정자 탈응축법의 적정화)

  • Pang, Myung-Geol
    • Clinical and Experimental Reproductive Medicine
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    • v.24 no.3
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    • pp.369-375
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    • 1997
  • Studies were conducted to determine the efficiency of decondensation protocols. Sperm obtained from seven normal donors was immediately washed after liquefaction and then decondensed using the method of West et al. (1989) and my original protocol. My optimized protocol entailed mixing 1 ml aliquots of semen with 4 ml phosphate buffered saline (PBS). Following centrifugation, pellets were resuspended in 1 ml PBS containing 6 mM EDTA. After centrifugation, pellets were resuspended in 1 ml PBS containing 2 mM dithiothreitol at $37^{\circ}C$ for 45 min. Following mixing with 2 ml PBS and centrifugation, pellets were resuspended by vortexing. While vortexing, 5 ml of fixative were gently added. Slide preparation was accomplished using the smear method and it was stored at $4^{\circ}C$. When comparing these protocols, the degree of sperm decondensation and head swelling was monitored by measuring nuclear length, area, perimeter, and degree of roundness using FISH analysis software. Apparent copy number for chromosome 1 and, separately, for the sex chromosomes was determined by FISH using satellite DNA probes for loci DIZ1, DXZ1 and DYZ3. Sperm treated by my decondensation protocol showed significant increases (p<0.05) in length, area, perimeter, and degree of roundness. There was a significant decrease (p<0.05) in the frequency of nuclei displaying no signal but no change in the frequency of nuclei with two signals in samples decondensed by my protocol. My data suggested that decondensation using my original protocol may lower the frequency of cells with spurious "nullisomy" due to hybridization failure without inducing spurious "disomy" resulting from increased distances between split signals.

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Development of Human Papillomavirus DNA Array by Using Lateral Flow Membrane Assay (Lateral Flow Membrane를 이용한 인유두종 바이러스 DNA Array의 개발)

  • Kim, Ki-Whang;Lee, Hyung-Ku;Cho, Hong-Bum
    • Korean Journal of Microbiology
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    • v.44 no.4
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    • pp.346-351
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    • 2008
  • This study develops DNA array which can detect specific sequence of human papilomavirus (HPV) by using lateral flow membrane assay which is usually used for point of care test including pregnant diagnosis. Principle of HPV DNA array is as follow; fixing DNA probe which is peculiar to HPV type 6, 11, 16, 18, 31, 45 on a surface of lateral flow membrane and inducing hybridization response between probe and HPV PCR products which is obtained by using biotin-labeled MY09/l1 primers. And then, we can see the result of DNA hybridization that streptavidin labelled colloidal gold is responded with hybrid biotin. Lateral flow membrane array developed in this study confirms major HPV type economically and conveniently compared with existing HPV DNA chip method.

Cloning of a Potentially Strain-Specific DNA Probe of prevotella intermedia ATCC 25611 by Inverted Dot Blot Hybridization Screening Method

  • Kook, Joong-Ki;Han, Jin-Ju;Kim, Hwa-Sook;Seong, Jin-Hyo;Kim, Dong-Kie;Baek, Dong-Heon;Choe, Son-Jin
    • Journal of Microbiology and Biotechnology
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    • v.13 no.2
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    • pp.282-286
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    • 2003
  • The purpose of this study was to isolate a specific DNA probe for the strain ATTC 25611 of the species Prevotella intermedia by using a new rapid screening mothod. The whole-genomic DNA of P. intermedia ATCC 25611 was isolated and purified. The HindIII-digested genomic DNAs from the strain were cloned by the random cloning method. To screen the strain-specific DNA probe, inverted dot blot hybridization tests were performed. In this assay, 20 ng of recombinant plasmids containing the HindIII-digested genomic DNA fragment were boiled and blotted onto a nylon membrane, and hybridized with digoxigenin-dUTP labeled genomic DNAs in a concentration of 100 ng/ml. Southern blot analysis was performed in order to confirm the results of the inverted dot blot hybridization tests. The data showed that a Pi34 probe (2.1 kbp; 1 out of 32 probes) was specific for P. intermedia strain ATCC 25611 and could be useful for the detection and identification of the strain, particularly in epidemiological studies of periodontal disease.