• Title/Summary/Keyword: Hybridization

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Studies on In Situ Hybridization of Electron Microscopy for Detection of Viral RNA (전자현미경 In Situ Hybridization에 의한 Viral RNA의 진단에 관한 연구)

  • 최원기;주경웅;김석홍
    • Biomedical Science Letters
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    • v.2 no.2
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    • pp.257-265
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    • 1996
  • Simple stain and electron microscopic in situ hybridization is studied and applied for the identification of rabbit haemorrhagic disease viral RNA in a unicrylated preparation of the liver after innoculation of rabbit haemorrhagic disease virus. Hybridization for detection of viral RNA in unicryl embedded tissues using complementary 84 bases oligonucleotide probe labelled by biotin CE-phosphoramidite compared with 4717∼4800 sequences of rabbit haemorrhagic disease virus, modified hybridization protocol and antibiotin antibody-l0nm gold as signal marker. The best results were obtained in 0.02% glutaraldehyde, Unicryl resin cell block, biotinylated oligonucleotide probes, antibiotin-l0nm gold. In this report, RHD viral RNA was distributed widely within the mitochondria and nucleus of liver cell by electron microscopic in situ hybridization. In situ hybridization has become a standard method for localizing DNA or RNA sequences in tissue or celt preparation. In situ hybridization is detected the virus genome in the cells and tissue as specifically compared with others nucleic acid hybridization method.

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Enhancement of DNA Microarray Hybridization using Microfluidic Biochip (미세유체 바이오칩을 이용한 DNA 마이크로어레이 Hybridization 향상)

  • Lee, H.H.;Kim, Y.S.
    • KSBB Journal
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    • v.22 no.6
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    • pp.387-392
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    • 2007
  • Recently, microfluidic biochips for DNA microarray are providing a number of advantages such as, reduction in reagent volume, high-throughput parallel sample screening, automation of processing, and reduction in hybridization time. Particularly, the enhancement of target probe hybridization by decrease of hybridization time is an important aspect highlighting the advantage of microfluidic DNA microarray platform. Fundamental issues to overcome extremely slow diffusion-limited hybridization are based on physical, electrical or fluidic dynamical mixing technology. So far, there have been some reports on the enhancement of the hybridization with the microfluidic platforms. In this review, their principle, performance, and outreaching of the technology are overviewed and discussed for the implementation into many bio-applications.

Detection and Analysis of DNA Hybridization Characteristics by using Thermodynamic Method (열역학법을 이용한 DNA hybridization 특성 검출 및 해석)

  • Kim, Do-Gyun;Gwon, Yeong-Su
    • The Transactions of the Korean Institute of Electrical Engineers C
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    • v.51 no.6
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    • pp.265-270
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    • 2002
  • The determination of DNA hybridization reaction can apply the molecular biology research, clinic diagnostics, bioengineering, environment monitoring, food science and application area. So, the improvement of DNA hybridization detection method is very important for the determination of this hybridization reaction. Several molecular biological techniques require accurate predictions of matched versus mismatched hybridization thermodynamics, such as PCR, sequencing by hybridization, gene diagnostics and antisense oligonucleotide probes. In addition, recent developments of oligonucleotide chip arrays as means for biochemical assays and DNA sequencing requires accurate knowledge of hybridization thermodynamics and population ratios at matched and mismatched target sites. In this study, we report the characteristics of the probe and matched, mismatched target oligonucleotide hybridization reaction using thermodynamic method. Thermodynamic of 5 oligonucleotides with central and terminal mismatch sequences were obtained by measured UV-absorbance as a function of temperature. The data show that the nearest-neighbor base-pair model is adequate for predicting thermodynamics of oligonucleotides with average deviations for $\Delta$H$^{0}$ , $\Delta$S$^{0}$ , $\Delta$G$_{37}$ $^{0}$ and T$_{m}$, respectively.>$^{0}$ and T$_{m}$, respectively.

Comparative Quantitative Study of Surfactant Protein C mRNA by Filter Hybridization and Solution Hybridization in Rats (Filter Hybridization과 Solution Hybridization 방법에 의한 백서 Surfactant Protein C mRNA 정량측정의 비교)

  • Kim, Jin-Ho;Sohn, Jang-Won;Yang, Seok-Chul;Yoon, Ho-Joo;Shin, Dong-Ho;Park, Sung-Soo
    • Tuberculosis and Respiratory Diseases
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    • v.51 no.6
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    • pp.517-529
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    • 2001
  • Background : Surfactant protein C(SP-C) is a hydrophobic 5,000 dalton molecule. SP-C has the primary roles in accelerating surface spreading of a surfactant phospholipid. The filter hybridization and solution hybridization assays are both rapid and sensitive and can be used to measure the RNAs complementary to any cloned DNA sequence. Methods : The authors measured the SP-C mRNA levels quantitatively using solution hybridization and filter hybridization assays to obtain a standard curve equation to quantify the mRNA of unknown samples comparatively. Results : 1. The minimum level of the specimens by solution hybridization was 3 pg for SP-C mRNA. 2. The standard curve equation of the solution hybridization assay between the counts per minute(Y) and the SP-C mRNA transcript input(X) was Y=6.46 X+244. The correlation coefficient was 0.99. 3. The minimum detection level of specimens by filter hybridization was 0.1 ng for SP-C mRNA. 4. The standard curve equation of the filter hybridization assay between the counts per minute(Y) and SP-C mRNA transcript input(X) is Y=2541.6 X+252.7. The correlation coefficient was 0.99. Conclusions : A comparison of CPM/filter in the linear range allowed an accurate and reproducible estimation of the SP-C mRNA copy number. Filter hybridization and solution hybridization assays are both rapid and sensitive and can be used to measure the RNAs complementary to any cloned DNA sequence. It is ideally suited to situations where accurate quantitation of multiple samples is required.

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A Study on Match and Mismatch DNA Hybridization properties Using DNA Hybridization Detection Sensor (DNA Hybridization 검출 센서를 이용한 매치 및 미스매치 DNA hybridization 특성 연구)

  • Kim, Do-Kyun;Kwon, Young-Soo
    • Proceedings of the KIEE Conference
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    • 2003.10a
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    • pp.89-91
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    • 2003
  • The determination of DNA hybridization reaction can apply the molecular biology research, clinic diagnostics, bioengineering, environment monitoring, food science and other application area. So, the improvement of DNA detection system is very important for the determination of this hybridization reaction. In this study, we report the characterization of the probe and target oligonucleotide hybridization reaction using the evanescent field microscopy. First, we have fabricated DNA chip microarray. The particles which were immobilized oligonucleotides were arranged by the random fluidic self-assembly on the pattern chips, using hydrophobic interaction. Second, we have detected DNA hybridization reaction using evanescent field microscopy. The 5'-biotinylated probe oligonucleotides were immobilized on the surface of DNA chip microarray and the hybridization reaction with the Rhodamine conjugated target oligonucleotide was excited fluorescence generated on the evanescent field microscopy. In the foundation of this result, we could be employed as the basis of a probe olidonucleotide, capable of detecting the target oligonucleotide and monitoring it in a large analyte concentration range and various mismatching condition.

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Comparison between Dot Blot Hybridization and Southern Blot Hybridization in Detecting Methicillin Resistant Staphylococcus aureus (Methicillin 내성 Staphylococcus aureus의 검출을 위한 분자유전학적 기법에 관한 연구)

  • 조태흠;김민정;오양효
    • Journal of Life Science
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    • v.9 no.4
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    • pp.358-367
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    • 1999
  • Thirty strains of methicillin resistant Staphylococcus aureus were obtained from the clinical isolates. In order to investigate the pursuit of the pathogens of nosocomial infection, these strains were studied for antibiotic sensitivity as well as its resistant pattern. Among the methods of hybridization which directly confirm the specific antibiotic resistant genes by means of the recently developed specific probe DNA, dot blot hybridization and southern blot hybridization were performed and these two methods were compared in their sensitivity and specificity. Strains that is sensitive to cephalothin to the subject of methicillin resistant Staphylococcus aureus were in 43%. Those that are sensitive to cefoperazone and cefuroxime were 26% and 23%, respectively. In case of MIC, MIC50 of cefoperazone was 8 $\mu\textrm{g}$/$m\ell$, and MIC90 was 128 $\mu\textrm{g}$/$m\ell$ to be the lowest. As the results of plasmid DNA electrophoresis, most of methicillin resistant Staphylococcus aureus strains had more than 4 plasmids. These plasmids digested by BamHI, methicillin resistant Staphylococcus aureus is distributed as 10 fragments with the size of 65 kb to 1.5 kb. Dot blot hybridization were performed to examine the existence of mecA gene to show the detection rate of 50%. Southern blot hybridization were done to see if DNA bands which amplify the activity of digoxigenium-labeled probe by PCR were actually PCR products of mecA gene and it showed the detection rate of 53%. It can be concluded that the southern blot hybridization seemed to be better in sensitivity and specificity when it is compared with the results of dot blot hybridization.

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Hybridization in Digital Geometry (디지털 조형의 혼성적 특성연구)

  • Kim, Ran-Hee;Ahn, Seongmo
    • Korean Institute of Interior Design Journal
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    • v.23 no.4
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    • pp.129-139
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    • 2014
  • The objective of this research is to suggest new geometric possibilities in digital architecture by investigating the characteristics of hybridization in digital geometry. The research begins with theoretical background research such as defining hybridization, investigating hybrid thinking, and studying the theory of digital geometry, along with the four conceptual characteristics of hybridization that could be drawn, such as temporality, liquidity, complexity, and connectivity. Based on these characteristics, the generative method of hybrid digital geometric languages such as Blob, Particle, Morph, Loft, and Boolean was analyzed with case research in contemporary digital architecture. As a result, diverse hybrid geometric keywords were extracted; these keywords suggest potential meanings of hybridization such as accidentality, mobility, diversity, and identity. Different elements represent the "mobility" in time by the force and wave, and they are "accidentally" combined in gradual change. The united species in "diverse" characters are seamlessly connected and emerge as a new "identity." The research maximizes the generative possibilities in digital geometry and provides a theoretical basis to apply the digital hybrid methods to architectural design by suggesting the potential meanings and possibilities in hybridization.

Directed Colony Hybridization Using Agarose Gel

  • Park, Jong-Chun;Chun, Soon-Bai
    • Journal of Microbiology and Biotechnology
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    • v.4 no.3
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    • pp.235-236
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    • 1994
  • Direct colony hybridization on agarose gel plate was established for the identification of recombinant plasmids. The hybridization of the probe to nucleic acids on dried gel without transferring to solid supports was more effective and simpler than hybridization of such probes to materials immobilized on filters such as nitrocellulose or nylon. D-cycloserine in overlaying agamse was essential for releasing the nucleic acids from colony.

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Relationship Between Genome Similarity and DNA-DNA Hybridization Among Closely Related Bacteria

  • Kang, Cheol-Hee;Nam, Young-Do;Chung, Won-Hyong;Quan, Zhe-Xue;Park, Yong-Ha;Park, Soo-Je;Desmone, Racheal;Wan, Xiu-Feng;Rhee, Sung-Keun
    • Journal of Microbiology and Biotechnology
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    • v.17 no.6
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    • pp.945-951
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    • 2007
  • DNA-DNA hybridization has been established as an important technology in bacterial species taxonomy and phylogenetic analysis. In this study, we analyzed how the efficiency with which the genomic DNA from one species hybridizes to the genomic DNA of another species (DNA-DNA hybridization) in microarray analysis relates to the similarity between two genomes. We found that the predicted DNA-DNA hybridization based on genome sequence similarity correlated well with the experimentally determined microarray hybridization. Between closely related strains, significant numbers of highly divergent genes (>55% identity) and/or the accumulation of mismatches between conserved genes lowered the DNA-DNA hybridization signal, and this reduced the hybridization signals to below 70% for even bacterial strains with over 97% 16S rRNA gene identity. In addition, our results also suggest that a DNA-DNA hybridization signal intensity of over 40% indicates that two genomes at least shared 30% conserved genes (>60% gene identity). This study may expand our knowledge of DNA-DNA hybridization based on genomic sequence similarity comparison and further provide insights for bacterial phylogeny analyses.

Interspecific Hybridization of the Bumblebees Bombus ignitus and B. terrestris

  • Yoon, Hyung-Joo;Kim, Seong-Yeol;Lee, Kyeong-Yong;Lee, Sang-Beom;Park, In-Gyun;Kim, Ik-Soo
    • International Journal of Industrial Entomology and Biomaterials
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    • v.18 no.1
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    • pp.41-48
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    • 2009
  • The large bumblebee, Bombus terrestris, indigenous to Europe and used extensively for high-value crop pollination, has been artificially introduced in several parts of the world. Here we show the interspecific hybridization between bumblebee species, B. terrestris and B. ignitus, under laboratory conditions. The mating and oviposition percentages of the interspecific hybridization of a B. terrestris queen with a B. ignitus male were higher than those of the intraspecific mating of B. ignitus. Furthermore, the competitive copulation experiment indicated that the mating of B. ignitus males with B. terrestris queens was 1.8-fold more frequent than with B. ignitus queens. The interspecific hybridization of a B. ignitus queen with a B. terrestris male produced either B. ignitus workers or the B. ignitus male phenotype, and the hybridization of a B. terrestris queen with a B. ignitus male produced B. terrestris males. Genetic tests using a portion of the mitochondrial COI gene for the parent and hybrid phenotypes indicated that mitochondrial DNA in the interspecific hybridization was maternally inherited. Our results indicated that interspecific hybridization occurred between B. ignitus and B. terrestris, which suggests that the hybridization will have a negative impact of competition and genetic pollution of native bumblebees.