• Title/Summary/Keyword: Hoechst dye

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Genome Detection Using Hoechst 33258 Groove Binder (Hoechst 33258 Groove Binder를 이용한 DNA칩)

  • Choi, Yong-Sung;Lee, Kyung-Sup
    • Proceedings of the Korean Institute of Electrical and Electronic Material Engineers Conference
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    • 2006.06a
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    • pp.372-373
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    • 2006
  • In this paper, a DNA chip with a microelectrode array was fabricated using microfabrication technology. Several probe DNAs consisting of mercaptohexyl moiety at their 5 end were immobilized on the gold electrodes by DNA arrayer. Then target DNAs were hybridized and reacted with Hoechst 33258, which is a DNA minor groove binder and electrochemically active dye. Linear sweep voltammetry or cyclic voltammetry showed a difference between target DNA and control DNA in the anodic peak current values. It was derived from Hoechst 33258 concentrated at the electrode surface through association with formed hybrid. It suggested that this DNA chip could recognize the sequence specific genes.

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Electrochemical Gene Detection Using Hoechat Groove Binder (Hoechst groove binder를 이용한 유전자의 전기화학적 검출)

  • Choi, Yong-Sung;Lee, Woo-Ki;Lee, Kyung-Sup
    • Proceedings of the Korean Institute of Electrical and Electronic Material Engineers Conference
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    • 2006.05a
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    • pp.65-70
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    • 2006
  • In this study, a DNA chip with a microelectrode array was fabricated using microfabrication technology. Several probe DNAs consisting of mercaptohexyl moiety at their 5 end were immobilized on the gold electrodes by DNA arrayer. Then target DNAs were hybridized and reacted with Hoechst 33258, which is a DNA minor groove binder and electrochemically active dye. Linear sweep voltammetry or cyclic voltammetry showed a difference between target DNA and control DNA in the anodic peak current values. It was derived from Hoechst 33258 concentrated at the electrode surface through association with formed hybrid. It suggested that this DNA chip could recognize the sequence specific genes.

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Circular Plasmid DNA from a Red Algae, Porphyra tenera (양식 참김(Porphyra tenera)에서 분리한 Circular Plasmid DNA)

  • 류태형;최학선;최경희;이춘환
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.27 no.6
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    • pp.1160-1165
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    • 1998
  • When total cellular DNA was isolated from Porphyra tenera by ultracentrifugation on Hoechst dye/CsCl gradients method, plasmid like DNA's were concentrated at the upper band which were characterized with a A+T rich organelle DNA's in the CsCl gradients. Based on their electrophoretic migration in different concentration of agarose gel, buffer system, and electric power etc. and the results of restriction digestion, the plasmid like DNA's were concluded to have circular conformation. This is the first report of putative circular plasmid DNA from the P. tenera, which is a autonomously replicating plasmid existing with a high copy number plasmid in the cell. The minimum size of this plasmid estimated by restriction endonuclease digestion was appeared to be 2.5kb in size.

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Electrochemical Gene Detection Using Microelectrode Array on a DNA Chip

  • Park, Yong-Sung;Kwon, Young-Soo;Park, Dae-Hee
    • KIEE International Transactions on Electrophysics and Applications
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    • v.4C no.4
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    • pp.145-148
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    • 2004
  • In this study, a DNA chip with a microelectrode array was fabricated using microfabrication technology. Several probe DNAs consisting of mercaptohexyl moiety at their 5' end were immobilized on the gold electrodes by a DNA arrayer. Then target DNAs were hybridized and reacted with Hoechst 33258, which is a DNA minor groove binder and electrochemically active dye. Linear sweep voltammetry or cyclic voltammetry showed a difference between target DNA and control DNA in the anodic peak current values. It was derived from Hoechst 33258 and concentrated at the electrode surface through association with the formed hybrid. This suggested that this DNA chip could recognize the sequence specific genes.

Eletrochemical Detection of Gene using Microelectrode-array DNA Chip (미소전극어레이형 DNA칩을 이용한 유전자의 전기화학적 검출)

  • ;;Eiichi Tamiya
    • Journal of the Korean Institute of Electrical and Electronic Material Engineers
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    • v.17 no.7
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    • pp.729-737
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    • 2004
  • In this paper, a DNA chip with a microelectrode array was fabricated using microfabrication technology. Several probe DNAs consisting of mercaptohexyl moiety at their 5 end were immobilized on the gold electrodes by DNA arrayer. Then target DNAs were hybridized and reacted with Hoechst 33258, which is a DNA minor groove binder and electrochemically active dye. Linear sweep voltammetry or cyclic voltammetry showed a difference between target DNA and control DNA in the anodic peak current values. It was derived from Hoechst 33258 concentrated at the electrode surface through association with formed hybrid. It suggested that this DNA chip could recognize the sequence specific genes.

Simple Classification of Male Mouse Germ Cells using Hoechst 33258 Staining (Hoechst 33258 Staining을 이용한 웅성 생쥐 성세포의 간편 분류)

  • Kim, Kyoung Guk;Park, Young Sik
    • Journal of Embryo Transfer
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    • v.30 no.3
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    • pp.213-218
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    • 2015
  • In the study for a differentiation and development of spermatogonial cells, the researchers should commonly require a simple, fast and reasonable method that could evaluate the developmental stage of male germ cells without any damage and also relentlessly culture them so far as a cell stage aiming at experimental applications. For developing the efficient method to identify the stage of sperm cells, the morphological characteristics of sperm cells were investigated by staining the cells with blue fluorescent dye Hoechst 33258, and a criterion for male germ cell classification was elicited from results of the previous investigation, then the efficiency of the criterion was verified by applying it to assort the germ cells recovered from male mice in age from 6 to 35 days. As morphological characteristics, spermatogonia significantly differed from spermatocytes in size, appearance and fluorescent patches of nucleus, and spermatids could also be distinguished from spermatozoa by making a difference in the volume and shape of nucleus and the shape and fluorescence of tail. Aforesaid criterion was applicable for classifying in vitro cultured sperm cells by verifying its efficiency and propriety for assorting the stages of testicular germ cells. However, the fluorescent staining showed that germ cells in mouse testis should be dramatically differentiated and developed at 21 days and 35 days of age, which were known as times of sexual puberty and maturity in male mice, respectively. In conclusion, the results indicated that this simple criterion for sperm cell classification using fluorescence staining with Hoechst 33258 may be highly efficient and reasonable for spermatogenesis study.

Genomic Detection using Electrochemical Method (전기화학적 방법에 의한 유전자의 검출)

  • Choi, Yong-Sung;Lee, Kyung-Sup;Park, Dae-Hee
    • Journal of the Korean Institute of Electrical and Electronic Material Engineers
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    • v.18 no.6
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    • pp.560-570
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    • 2005
  • In this paper, a microelectrode away DNA chip was fabricated on glass slide using photolithography technology. Several probe DNAs consisting of mercaptohexyl moiety at their 5' end were immobilized on the gold electrodes by DNA arrayer utilizing the affinity between gold and sulfu. Then target DNAs were hybridized and reacted with Hoechst 33258, which is a DNA minor groove binder and electrochemically active dye. Cyclic voltammetry in 5mA ferricyanide/ferrocyanide solution at 100 mV/s confirmed the immobilization of probe DNA on the gold electrodes. Linear sweep voltammetry or cyclic voltammetry showed a difference between target DNA and control DNA in the anodic peak current values. It was derived from Hoechst 33258 concentrated at the electrode surface through association with formed hybrid. It suggested that this DNA chip could recognize the sequence specific genes. It suggested that multichannel electrochemical DNA microarray is useful to develop a portable device for clinical gene diagnostic system.

Electrochemical Detection of Genes Using Microeledtrode Array DNA Chip (미소전극어에이형 DNA칩을 이용한 유전자의 전기화학적 검출)

  • Choi, Yong-Sung;Park, Dae-Hee
    • Proceedings of the KIEE Conference
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    • 2004.07c
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    • pp.2125-2127
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    • 2004
  • In this paper, a DNA chip with a microelectrode array was fabricated using microfabrication technology. Several probe DNAs consisting of mercaptohexyl moiety at their 5 end were immobilized on the gold electrodes by DNA arrayer. Then target DNAs were hybridized and reacted with Hoechst 33258, which is a DNA minor groove binder and electrochemically active dye. Linear sweep voltammetry or cyclic voltammetry showed a difference between target DNA and control DNA in the anodic peak current values. It was derived from Hoechst 33258 concentrated at the electrode surface through association with formed hybrid. It suggested that this DNA chip could recognize the sequence specific genes.

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Side Population Cell Level in Human Breast Cancer and Factors Related to Disease-free Survival

  • Jin, C.G.;Zou, T.N.;Li, J.;Chen, X.Q.;Liu, X.;Wang, Y.Y.;Wang, X.;Che, Y.H.;Wang, X.C.;Sriplung, Hutcha
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.3
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    • pp.991-996
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    • 2015
  • Side population (SP) cells have stem cell-like properties with a capacity for self-renewal and are resistant to chemotherapy and radiotherapy. Therefore the presence of SP cells in human breast cancer probably has prognostic value. Objective: To investigate the characteristics of SP cells and identify the relationship between the SP cells levels and clinico-pathological parameters of the breast tumor and disease-free survival (DFS) in breast cancer patients. Materials and Methods: A total of 122 eligible breast cancer patients were consecutively recruited from January 1, 2006 to December 31, 2007 at Yunnan Tumor Hospital. All eligible subjects received conventional treatment and were followed up for seven years. Predictors of recurrence and/or metastasis and DFS were analyzed using Cox regression analysis. Human breast cancer cells were also obtained from fresh human breast cancer tissue and cultured by the nucleic acid dye Hoechst33342 with Verapami. Flow cytometry (FCM) was employed to isolate the cells of SP and non-SP types. Results: In this study, SP cells were identified using flow cytometric analysis with Hoechst 33342 dye efflux. Adjusted for age, tumor size, lymph nodal status, histological grade, the Cox model showed a higher risk of recurrence and/or metastasis positively associated with the SP cell level (1.75, 1.02-2.98), as well as with axillary lymph node metastasis (2.99, 1.76-5.09), pathology invasiveness type (1.7, 1.14-2.55), and tumor volume doubling time (TVDT) (1.54, 1.01-2.36). Conclusions: The SP cell level is independently associated with tumor progression and clinical outcome after controlling for other pathological factors. The axillary lymph node status, TVDT and the status of non-invasive or invasive tumor independently predict the prognosis of breast cancer.

Zinc-induced Apoptosis in C6 glial Cells via Generation of Hydrogen Peroxide($H_2O_2$) (신경교세포주 C6 glial에서 Zinc의 Hydrogen Peroxide($H_2O_2$) 생성을 통한 세포고사)

  • 이지현;김명선;소흥섭;김남송;조광호;이향주;이기남;박길래
    • Toxicological Research
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    • v.16 no.3
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    • pp.179-185
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    • 2000
  • Zinc is known to generate reactive oxygen species (ROS) including superoxide anion and hydrogen peroxide ($H_2O_2$), which eventually contribute to cytotoxicity in a variety of cell types. Here in, we demonstrated that zinc decreased the viability of C6 glial cells in a time and dose-dependent manner, which was revealed as apoptosis characterized by ladder-pattern fragmentation of genomic DNA. chromatin condensation and DNA fragmentation in Hoechst dye staining. Zinc-induced apoptosis of C6 glial cells was prevented by the addition of catalase and antioxidants including reduced glutathione (GSH), N-acetyl-L-cysteine (NAC) and pyrrolidinedithiocarbamate (PDTC). Wefurther confirmed that zinc decreased intrac-ellular levels of GSH and generated $H_2O_2$in C6 glial cells. Moreover, antioxidants also decreased the generation of zinc-induced $H_2O_2$ in C6 glial cells. These data indicated that zinc-induced the apoptotic death of C6 glial cells via generation of reactive oxygen species such as $H_2O_2$.

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