• Title/Summary/Keyword: DNA recovery

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DNA Microarray Probe Preparation by Gel Isolation Nested PCR

  • Wang, Hong-Min;Ma, Wen-li;Huang, Hai;Xiao, Wei-Wei;Wang, Yan;Zheng, Wen-Ling
    • BMB Reports
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    • v.37 no.3
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    • pp.356-361
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    • 2004
  • To develop a simplified method that can rapidly prepare DNA microarray probes in a massive scale, a lambda phage genomic DNA-fragments library was constructed for the microarray-probes collection. Four methods of DNA band recovery from the first PCR products were tested and compared. The DNA microarray probes were collected by a novel method of nested PCR that was mediated by gel isolation of the first PCR products. This method was named GIN-PCR. The probes that were prepared by this GIN-PCR technique were used as subjects to fabricate a DNA microarray. The results showed that a wooden toothpick was superior to the other 3 methods, since this technique can steadily transfer the DNA bands as the template of the second PCR after the first PCR. A group of probes were successfully collected and DNA microarrays were constructed using these probes. Hybridization results demonstrated that this technique of DNA recovery and probe preparation was rapid, efficient, and effective. We developed a cost-effective and less labor-intensive method for DNA microarray probe preparation by nested PCR that is mediated by wooden toothpick transfer of the DNA bands in the gel after electrophoresis.

DNA Double-Strand Breaks Serve as a Major Factor for the Expression of Arabidopsis Argonaute 2

  • Lee, Sungbeom;Chung, Moon-Soo;Lee, Gun Woong;Chung, Byung Yeoup
    • Journal of Radiation Industry
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    • v.10 no.4
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    • pp.243-248
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    • 2016
  • Argonaute 2 (AtAGO2) is a well characterized effector protein in Arabidopsis for its functionalities associated with DNA double-strand break (DSB)-induced small RNAs (diRNAs) and for its inducible expression upon ${\gamma}$-irradiation. However, its transcriptional regulation depending on the recovery time after the irradiation and on the specific response to DSBs has been poorly understood. We analyzed the 1,313 bp promoter sequence of the AtAGO2 gene ($1.3kb_{pro}$) to characterize the transcriptional regulation of AtAGO2 at various recovery times after ${\gamma}$-irradiation. A stable transformant harboring $1.3kb_{pro}$ fused with GUS gene showed that the AtAGO2 is highly expressed in response to ${\gamma}$-irradiation, after which the expression of the gene is gradually decreased until 5 days of DNA damage recovery. We also confirm that the AtAGO2 expression patterns are similar to that of ${\gamma}$-irradiation after the treatments of radiomimetic genotoxins (bleomycin and zeocin). However, methyl methanesulfonate and mitomycin C, which are associated with the inhibition of DNA replication, do not induce the expression of the AtAGO2, suggesting that the expression of the AtAGO2 is closely related with DNA DSBs rather than DNA replication.

An research of the error detection method and efficient recovery algorithms in the DNA double helix. (DNA 이중나선에서의 오류위치 검출 방법 및 효율적인 복구 알고리즘 연구)

  • Kim, Soke-Hwan;Hur, Chang-Wu
    • Proceedings of the Korean Institute of Information and Commucation Sciences Conference
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    • 2012.10a
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    • pp.293-297
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    • 2012
  • In order to maintain order in the genetic information at cells, it need ongoing monitoring and recovery system. DNA is accomplished by a combination of base pairs, Wrong base pairs is formed with a much more lower frequency than the normal DNA. if it does not modify and was accumulate, the Cells were died. In this study, mistakes of DNA replication and repair of the damaged part was introduced engineering concepts by mimicking DNA repair functions. It was presented recover the complementary part of the previously announced and presented an efficient algorithm at find and recover the complementary part.

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An research of the error detection method and efficient recovery algorithms in the DNA double helix (DNA 이중나선에서의 오류위치 검출 방법 및 효율적인 복구 알고리즘 연구)

  • Kim, SokeHwan;Hur, Chang-Wu
    • Journal of the Korea Institute of Information and Communication Engineering
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    • v.16 no.11
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    • pp.2557-2562
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    • 2012
  • In order to maintain order in the genetic information at cells, it need ongoing monitoring and recovery system. DNA is accomplished by a combination of base pairs, Wrong base pairs is formed with a much more lower frequency than the normal DNA. if it does not modify and was accumulate, the Cells were died. In this study, mistakes of DNA replication and repair of the damaged part was introduced engineering concepts by mimicking DNA repair functions. It was presented recover the complementary part of the previously announced and presented an efficient algorithm at find and recover the complementary part.

Evidence for the Ras-Independent Signaling Pathway Regulating Insulin-Induced DNA Synthesis

  • Jhun, Byung-H.
    • BMB Reports
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    • v.32 no.2
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    • pp.196-202
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    • 1999
  • The existence of the Ras-independent signal transduction pathway of insulin leading to DNA synthesis was investigated in Rat-1 fibroblasts overexpressing human insulin receptor (HIRc-B) using the single-cell microinjection technique. Microinjection of a dominant-negative mutant $Ras^{N17}$ protein into quiescent HIRc-B cells inhibited the DNA synthesis stimulated by insulin. Microinjection of oncogenic H-$Ras^{V12}$ protein ($H-Ras^{V12}$) (0.1 mg/ml) induced DNA synthesis by 35%, whereas that of control-injected IgG was induced by 20%. When the marginal amount of oncogenic H-$Ras^{V12}$ protein was coinjected with a dominant-negative mutant of the H-Ras protein ($Ras^{N17}$), DNA synthesis was 35% and 74% in the absence and presence of insulin, respectively. This full recovery of DNA synthesis by insulin suggests the existence of the Ras-independent pathway. The same recovery was observed in the cells coinjected with either H-$Ras^{V12}$ plus H-$Ras^{N17}$ plus SH2 domain of the p85 subunit of PI3-kinase ($p85^{SH2-N}$) or H-$Ras^{V12}$ plus H-$Ras^{N17}$ plus interfering anti-Shc antibody. When co-injected with a dominant-negative H-$Ras^{N17}$, the DNA synthesis induced by the Ras-independent pathway was blocked. These results indicate that the Ras-independent pathway of insulin leading to DNA synthesis exists, bypassing the p85 of PI3-kinase and Shc protein, and requires Rac1 protein.

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Vapor Detection of ssDNA Decorated Graphene Transistor (ssDNA를 이용한 그래핀 가스 센서)

  • Jung, Youngmo;Kim, Young Jun;Moon, Hi Gue;Kim, Soo Min;Shin, Beomju;Lee, Joo Song;Seo, Minah;Lee, Taikjin;Kim, Jae Hun;Jun, Seong Chan;Lee, Seok;Kim, Chulki
    • Journal of Sensor Science and Technology
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    • v.23 no.5
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    • pp.310-313
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    • 2014
  • We report a way to improve the ability of graphene to operate as a gas sensor by applying single stranded deoxyribonucleic acid (DNA). The sensitivity and recovery of the DNA-graphene sensor depending on the different DNA sequences are analyzed. The different sensor responses to reactive chemical vapors are demonstrated in the time domain. Because of the chemical gating effect of the deposited DNA, the resulting devices show complete and rapid recovery to baseline unlike the bare graphene at room temperature. The application of the pattern recognition technique can increase the potential of DNA-graphene sensors as a chemical vapor classifier.

The Effects of Chungganhaeju-tang(Qingganjiejiu-tang) on Ethanol-mediated Cytokine Expression (청간해주탕이 에탄올 매개성 cytokine 발현에 미치는 영향)

  • 김병삼;김영철;이장훈;우홍정
    • The Journal of Korean Medicine
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    • v.24 no.1
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    • pp.190-201
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    • 2003
  • Object : This study was designed to investigate the effects of Chungganhaeju-tang (Qingganjiejiu-tang) on cytotoxicity, growth inhibition, apoptosis and expression of cytokine in damaged HepG2 cells. Method : Toxicity on HepG2 cell induced by ethanol and acetaldehyde was measured for viability, cell growth, DNA replication and generation of apoptosis and cytokine. The recovery of the cell activity by Chungganhaeju-tang was estimated for the measured parameters using PCR with different cycle numbers, DNA gel-electrophoresis, and densitometric analysis, Results : Chungganhaeju-tang improves the recovery of HepG2 cells damaged by ethanol or acetaldehyde. The suppressed DNA synthesis of the cell damaged by ethanol or acetaldehyde is improved by Chungganhaeju-tang. A liver-protection effect was shown by the reduction of apoptosis and $TNF-{\alpha},{\;}IL-1{\beta}$ expressions that are induced by ethanol or acetaldehyde. Conclusion : The result indicates that Chungganhaeju-tang reduces toxicity induced by ethanol or acetaldehyde and recovers damaged liver function.

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Improvement of the electro-transformed cell yield for Bifidobacterium sp. with large DNA

  • Park, Min Ju;Park, Myeong Soo;Ji, Geun Eog
    • Korean Journal of Food Science and Technology
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    • v.51 no.2
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    • pp.188-191
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    • 2019
  • In this study, the poor electro-transformant yield of large DNA in bifidobacteria was improved by increasing the DNA concentration, which was amplified by enhancing electroporation conditions: treating the cell wall weakening agent and cell membrane permeabilizing molecule as well as changing the electrical parameter. In the enhanced conditions, the electroporation frequency increased from 15 to 71 times compared to the initial conditions at the same DNA concentration. As the DNA concentration increased, the difference in the electroporation frequency between the two conditions became greater, and the curve of the enhanced conditions seemed to be saturated with a DNA concentration over $4{\mu}g$. The present study provided a clue to the recovery of the electroporation frequency with large DNA and formulated the relationship between the DNA concentration, the DNA size and the electroporation frequency in bifidobacteria. Therefore, this study will contribute to the expansion of molecular genetic studies of bifidobacteria.

Methods for the Extraction of DNA from Water Samples for Polymerase Chain Reaction

  • Jung, Jae-Sung;Lee, Young-Jong
    • Journal of Microbiology
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    • v.35 no.4
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    • pp.354-359
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    • 1997
  • Methods for the extraction of DNA from water sample were approximated. Four different procedures of DNA extraction were carried out with pellets obtained from centrifugation of 4 liter water samples. The recovery efficiency and purity of DNA extracted by each method from different sources were compared. DNA yield varied with extraction methods, Method I, which involves enzymatic and freeze-thaw lysis steps and phenol and phenol-chloroform purification of extracted nucleic acid, showed a significantly higher yield and purity than the other methods. The use of glass beads in the DNA extraction methods improved the purity of DNA suitable for PCR. Bovine serum albumin in the PCR reaction mixture was useful in reducing inhibitory effects of contaminants. The efficiency of an extraction method was determined by the detection of the aer of Aeromonas hydrophila with PCR. The lower limit of detection of A. hydrophila from seeded tap water was 2 CFU/ml in PCR when method I was used for DNA preparation.

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Effects of the NADPH Oxidase p22phox C242T Polymorphism on Endurance Exercise Performance and Oxidative DNA Damage in Response to Aerobic Exercise Training

  • Paik, Il-Young;Jin, Chan-Ho;Jin, Hwa-Eun;Kim, Young-Il;Cho, Su-Youn;Roh, Hee-Tae;Suh, Ah-Ram;Suh, Sang-Hoon
    • Molecules and Cells
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    • v.27 no.5
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    • pp.557-562
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    • 2009
  • We examined the effects of the NADPH oxidase p22phox C242T polymorphism on endurance exercise performance and oxidative DNA damage in response to acute and chronic exercises. One hundred three subjects were recruited, among which 26 healthy subjects (CC: 12, TC: 12, and TT: 2) were studied during rest, exercise at 85% $VO_2max$, and recovery before and after 8 weeks of treadmill running. Lymphocyte DNA damage increased significantly in response to exercise (p < 0.05). There were no significant differences in plasma MDA, SOD concentrations and lymphocyte DNA damage between CC genotype and T allele group, but significant endurance training differences were observed. Endurance training increased exercise time to exhaustion in both the CC genotype and T allele groups (p < 0.05) but no significant difference was found between groups. The results of the current study with young, healthy, Korean men are interpreted to mean that 1) the majority had the CC genotype of the NADPH oxidase p22phox C242T polymorphism (82.5%: CC, 15.5%: TC, 1.9%: TT), 2) acute exercise increased lymphocyte DNA damage, 3) endurance training significantly increased exercise time to exhaustion, and alleviated lymphocyte DNA damage, and 4) The NADPH oxidase p22phox C242T polymorphism, however, did not alter lymphocyte DNA damage or exercise performance at rest, immediately after exercise, or during recovery.