• Journal of Microbiology and Biotechnology
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• v.19 no.11
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• pp.1408-1414
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• 2009

The obtention of high yields of purified plasmid DNA is viewed as an essential issue to be considered towards efficient production of DNA vaccines and therapeutic plasmids. In this work, Escherichia coli $DH5\alpha$. bearing the pVAXI-LacZ plasmid was grown in a developed semi-defined medium at different temperatures and tryptone concentrations. Analysis of pDNA yields and E. coli morphology revealed that at higher temperatures (37 and $40^{\circ}C$), higher specific yields and E. coli filamentation were obtained. However, the best results were achieved when a lower tryptone concentration was used. This approach was shown to be a powerful tool to promote plasmid amplification, keeping the desirable plasmid structure, and favoring the attainment of quality. Our results suggest that by using tryptone alone as an amino acid source, pDNA amplification was improved and a specific yield of 20.43 mg pDNA/g dcw was achieved, proving that this strategy can improve pDNA yield even at a small scale.

• Biomedical Science Letters
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• v.15 no.3
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• pp.229-232
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• 2009

DNA isolation for PCR-based short tandem repeat (STR) analysis is essential to recover high yields of amplifiable DNA from low copy number (LCN) DNA samples. There are different methods developed for DNA extraction from the small bloodstain and gloves, commonly found at crime scenes. In order to obtain STR profiles from LCN DNA samples, DNA extraction protocols, namely the automated $iPrep^{TM}$ $ChargeSwitch^{(R)}$ method, the automated $QIAcube^{TM}$ method, the automated $Maxwell^{(R)}$ 16 DNA $IQ^{TM}$ Resin method, and the manual $QIAamp^{(R)}$ DNA Micro Kit method, were evaluated. Extracted DNA was quantified by the $Quantifiler^{TM}$ Human DNA Quantification Kit and DNA profiled by $AmpFISTR^{(R)}$ $Identifiler^{(R)}$ Kit. Results were compared based on the amount of DNA obtained and the completeness of the STR profiles produced. The automated $iPrep^{TM}$ $ChargeSwitch^{(R)}$ and $QIAcube^{TM}$ methoas produced reproducible DNA of sufficient quantity and quality trom the dried blood spot. This two automated methods showed a quantity and quality comparable to those of the forensic manual standard protocols normally used in our laboratory. In our hands, the automated DNA extraction method is another obvious choice when the forensic case sample available is bloodstain. The findings of this study indicate that the manual simple modified $QIAamp^{(R)}$ DNA Micro Kit method is best method to recover high yields of amplifiable DNA from the numerous potential sources of LCN DNA samples.

• Biomedical Science Letters
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• v.2 no.2
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• pp.241-247
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• 1996

Poor yields of amplified DNAs could be resulted in polymerase chain reaction(PCR) processes with G+C-rich DNA primers because of their high $T_m$ values. To maximize the yields of amplification in PCR processes with G+C-rich primers, we compared the yields of amplified DNA fragments according to the concentrations of specific denaturants added to the reaction mixture of PCR system. With addition of the mixture of 2.5% glycerol and 1.25% formamide, or 2.5% dimethyl sulfoxide to the reaction cocktail, respectively, remarkable increases in the yields of amplified DNA fragments were not observed in the PCR systems with G+C-low primers of Lyl chromosomal gene from Borrelia burgdorferi but observed in the PCR system with G+C- ich primers of Is900 gene from Mycobacterium parahberculosis. Although we were not practically able to discriminate the yields of PCR DNAs according to the concentrations used in this study, addition of the mixture of 5% glycerol and 2.5% formamide, or 5% DMSO tended to increase the production of extra bands.

• Journal of Plant Biology
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• v.16 no.1_2
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• pp.30-38
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• 1973

To study the response to plant growth by the environmental factors, the effects of application of nitrogen on changes in the yield, crude protein, amino acids, chlorophyll, carotene, total phosphorus, acid-soluble phosphorus, phospholipids, RNA, and DNA were investigated with westerworlds 9Lolium sublatum) and perennial rye-grasses (Lolium perenne). The amounts of dry weight, crude protein, amino acids, chlorophyll, carotene, total phosphorus, acid-soluble phosphorus, phospholipids, RNA and DNA of both rye-grasses increased with adequately increasing nitrogen, and reached a maximum with an adequate application of nitrogen. The relationships between yields and crude protein contents, crude protein and RNA contents, and yields and RNA contents of westerworlds and perennial rye-grasses were found to be positively correlated, respectively. Therefore, in general, the response to plant growth by the environmental factors such as nitrogen nutrient may be summarized as follows: Environmental factors\longrightarrowDNA\longrightarrowRNA\longrightarrowProtein\longrightarrowPlant growth

• Bulletin of the Korean Chemical Society
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• v.25 no.11
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• pp.1667-1670
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• 2004

In DNA microarray produced by DNA-deposition technology, DNA-immobilization and -hybridization yields on a solid support are most important factors for its accuracy and sensitivity. We have developed a dendrimeric support using silylated aldehyde slides and polyamidoamine (PAMAM) dendrimers. An oligonucleotide array was prepared through a crosslinking between the dendrimeric support and an oligonucleotide. Both DNAimmobilization and -hybridization yields on the solid support increased by the modification with the dendrimers. The increase of the immobilization and hybridization efficiency seems to result from a threedimensional arrangement of the attached oligonucleotide. Therefore, our dendrimeric support may provide a simple and efficient solution to the preparation of DNA microarrays with high-density DNA-deposition and high hybridization efficiency.

• Korean Journal of Microbiology
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• v.29 no.1
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• pp.25-33
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• 1991

Antiviral activities of papaverine and nucleoside analogs, 9-[(1,3-dihydroxy-2-propoxy)methyl]guanine (DHPG) and acyclovir, against human cytomegalovirus (HCMV) infection were compared in vitro. Papaverine and DHPG were effective in reducing infectious HCMV yields with $ED_{50}{\s}$ (effective dose 50: the concentraion at which 50% of virus yields was obtained) of approximately 1.02 and $0.45{\mu}{\M}$, respectively; while acyclovir was less effective with an $ED_{50}$ of about $10.4{\mu}{\M}$The relative cytotoxicity of these drugs was evaluated under the same conditions used to measure infectious HCMV yields. Papaverine and DHPG demonstrated little cellular toxicity as measured by their effect on the viability of confluent cells at concentrations in the range of those demonstrating potent inhibition of HCMV replication. Similarly, protein synthesis was largely unaffected by these drugs in stationary mock-infected cells as measured by the incorporation of isotopically labelled amino acids. In contrast, cellular DNA synthesis was invariably reduced in the presence of either drug. HCMV-specific DNA synthesis was also strongly inhibited by papaverine and DHPG.

• Proceedings of the Korean Vacuum Society Conference
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• 2016.02a
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• pp.140.1-140.1
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• 2016

We have measured DNA translocation through a nanocapillary functionalized with probe DNA. These DNA-functionalized nanocapillaries selectively facilitate the translocation of target ssDNAs that are complementary to the probe ssDNAs. In addition, translocation of the complementary target ssDNA exhibits two tendencies to translocation speed, such as fast and slow translocation, whereas that of non-complementary target ssDNA yields only one tendency, fast translocation. These observations suggest that the complementary and non-complementary target ssDNAs may be discriminated due to different interaction strengths between target and probe ssDNAs. The temperature dependence measurements of DNA translocation show that slow translocation events are ascribed to the complementary interaction between probe and target ssDNA. This confirms that their dwell time is dependent on the base-pair binding strength. These results demonstrate that mere single-base different target DNA can be selectively detectable by using the probe DNA-functionalized nanocapillaries.

• Journal of Microbiology and Biotechnology
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• v.14 no.1
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• pp.162-166
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• 2004

Phage P22 tailspike is a thermostable homotrimeric protein, and temperature-sensitive folding (tsf) and global suppressor mutations affect its folding yields at elevated temperatures. We earlier suggested that the folding of the tailspike protein in Escherichia coli requires an unidentified molecular chaperone. Accordingly, in the present study, the interactions of purified DnaK, DnaJ, and GrpE heat-shock proteins with the tailspike protein were investigated during the translation and folding of the protein. The cotranslational addition of DnaJ to the tailspike protein resulted in the arrest of folding, when Dnak and GrpE were missing. However, the presence of DnaK, DnaJ, and GrpE had no effect on the folding yield of the tails pike protein, thus, providing evidence for the binding of the nascent tailspike protein with DnaJ protein, a member of DnaK chaperoning cycle.

• Proceedings of the Materials Research Society of Korea Conference
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• 2009.11a
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• pp.12.2-12.2
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• 2009

The work describes anoptimized process to highly efficient and convenient preparation in highthroughput magnetic human DNA separation with chemically functionalizedsilica-coated magnetic nanoparticles. The effect of nanoparticle's size and the surface's hydrophilicity change were studied for magnetic DNA separation process, inwhich the optimum efficiency was explored via the function of the amino-groupnumbers, particle size, the amount of the nanoparticles used, and theconcentration of NaCl salt. The DNA adsorption yields were high in terms of theamount of triamino-functionalized nanoparticles used, and the average particlesize was 25 nm. The adsorption efficiency of aminofunctionalized nanoparticleswas the 4-5 times (80-100%) higher compared to silica-coated nanoparticles only(10-20%). DNA desorption efficiency showed an optimum level of over 0.7 M ofthe NaCl concentration. To elucidate the agglomeration of nanoparticles afterelectrostatic interaction, the Guinier plots were calculated from small angleX-ray diffractions in a comparison of the results of electron diffraction TEM,and confocal laser scanning microscopy. Additionally, the direct separation ofhuman genomic DNA was achieved from human saliva and whole blood with highefficiency.

• Biotechnology and Bioprocess Engineering:BBE
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• v.8 no.4
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• pp.263-267
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• 2003

Magnetic nanoparticles prepared from an alkaline solution of divalent and trivalent iron ions could covalently bind protein via the activation of Nethyl-N-(3-dimethylaminopropyl) carbodiimide (EDC). Trypsin and avidin were taken as the model proteins for the formation of protein-nanoparticle conjugates. The immobilized yield of protein increased with molar ratio of EDC/nanoparticie. Higher concentrations of added protein could yield higher immobilized protein densities on the particles. In contrast to EDC, the yields of protein immobilization via the a ctivation of cyanamide were relatively lower. Nanoparticles bound with avidin could attach a single-stranded DNA through the avidin-biotin interaction and hybridize with a DNA probe. The DNA hybridization was confirmed by fluorescence microscopy observations. Immobilized DNA on nanoparticles by this technique may have widespread applicability to the detection of specific nucleic acid sequence and targeting of DNA to particular cells.