• 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.

• Molecules and Cells
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• v.27 no.2
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• pp.205-209
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• 2009

The loci of the 5S and 45S rRNA genes were localized on chromosomes in five species of Capsicum, namely, annuum, chacoense, frutescens, baccatum, and chinense by FISH. The 5S rDNA was localized to the distal region of one chromosome in all species observed. The number of 45S rDNA loci varied among species; one in annuum, two in chacoense and frutescens, and chinense, and four in baccatum, with the exceptions that 'CM334' of annuum had three loci and 'tabasco' of frutescens gad one locus. 'CM334'-derived BAC clones, 384B09 and 365P05, were screened with 5S rDNA as a probe, and BACs 278M03 and 262A23 were screened with 25S rDNA as a probe. Both ends of these BAC clones were sequenced. FISH with these BAC probes on pachytenes from 'CM334' plant showed one 5S rDNA locus and three 45S rDNA loci, consistent with the patterns on the somatic chromosomes. The 5S rDNA probe was also applied on extended DNA fibers to reveal that its coverage measured as long as 0.439 Mb in the pepper genome. FISH techniques applied on somatic and meiotic chromosomes and fibers have been established for chili to provide valuable information about the copy number variation of 45S rDNA and the actual physical size of the 5S rDNA in chili.

• The Korean Journal of Zoology
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• v.18 no.3
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• pp.131-140
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• 1975

Dose response for the nuscheduled DNA synthesis induced by various concentration of MMS was dose dependent and directly proportional to dose increased. Time dependence of unscheduled DNA synthesis was continued up to 4 hours, with the peak appearing 2$\\sim$3 hours after treatment with MMS and $^3 H$-thymidine labeling. Single treatment with BUdR or IUdR does not induce unscheduled DNA synthesis. BUdR and IUdR greatly enhanced MMS-induced unscheduled DNA synthesis, but the dose responses were different from that of single treatment with MMS. IUdR was found to be more effective sensitizer on MMS-induced unscheduled DNA synthesis.

• Korean Journal of Medicinal Crop Science
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• v.12 no.6
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• pp.515-518
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• 2004

Anemarrhena asphodeloides, a medicinal plant, has chromosome number of 2n=2x=22. To characterize the somatic metaphase chromosomes, physical mapping of 45S and 5S rDNAs using McFISH (multi-color fluorescence in situ hybridization) was applied. Two pairs of 45S rDNA loci were detected on the terminal regions of the short arm of chromosomes 1 and 3. A pair of 5S rDNA signal was observed on the short arm of chromosome 3. 5S rDNA site seemed to be the same locus as one of the 45S rDNA site. McFISH was very useful tool for the localization and identification of rDNAs on the metaphase chromosomes in A. asphodeloides.

• Bulletin of the Korean Chemical Society
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• v.34 no.12
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• pp.3665-3670
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• 2013

The R3V6 peptide includes a hydrophilic arginine stretch and a hydrophobic valine stretch. In previous studies, the R3V6 peptide was evaluated as a gene carrier and was found to have low cytotoxicity. However, the transfection efficiency of R3V6 was lower than that of poly-L-lysine (PLL) in N2A neuroblastoma cells. In this study, the transfection efficiency of R3V6 was improved in combination with high mobility group box 1A domain (HMGA). HMGA is originated from the nuclear protein and has many positively-charged amino acids. Therefore, HMGA binds to DNA via charge interaction. In addition, HMGA has a nuclear localization signal peptide and may increase the delivery efficiency of DNA into the nucleus. The ternary complex with HMGA, R3V6, and DNA was prepared and evaluated as a gene carrier. First, the HMGA/DNA complex was prepared with a negative surface charge. Then, R3V6 was added to the complex to coat the negative charges of the HMGA/DNA complex, forming the ternary complex of HMGA, R3V6, and DNA. A physical characterization study showed that the ternary complex was more stable than the PLL/DNA complex. The HMGA/R3V6/DNA complex had a higher transfection efficiency than the PLL/DNA, HMGA/DNA, or R3V6/DNA complexes in N2A cells. Furthermore, the HMGA/R3V6/DNA complex was not toxic to cells. Therefore, the HMGA/R3V6/DNA complex may be a useful gene delivery carrier.

• The Korean Journal of Zoology
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• v.20 no.2
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• pp.93-99
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• 1977

Dose response forDNA repair synthesis induced by various concentrations of MMC (0.05 $\\sim$ 0.5 $\\mu$g/ml) in HeLa $S_3$ cells was not dose-dependent and the amounts of it were relatively lower, representing $7\\sim9%$ of total DNA synthesizing cells in $0.1\\sim0.5 \\mug/ml$ concentrations. Time dependence study showed that MMC-induced DNA repair synthesis occurred as long as for 24 hours with similar incidences in all time courses. Pretreatment with BUdR was found to have a sensitization effect on MMC-induced DNA repair synthesis, but that with IUdR was not. Combined treatment with BUdR of IUdR and MMC suppressed remarkably the semiconservative DNA synthesis especially at later time course. These results seem to suggest that damages induced in DNA by MMC might be repaired by both fast and slow excision processes.

• Microbiology and Biotechnology Letters
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• v.14 no.2
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• pp.133-137
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• 1986

17S-ribosomal RNA gene from the hygromycin resistant protozoan Tetrahymena thermophila hmr 3 was cloned on E. coli vector pBR 322 as part of study to work the 17S-rRNA structure and the mechanism of hygromycin resistance. The 17S-rDNA was inserted into the Hind 111 site of pBR 322. The clones having recombinant plasmid were selected by the method of colony hybridization with a 17S-rDNA probe of wild type B1868. The orientation of 17S-rDNA insert was located near the tetracycline resistant gene of pBR 322 in a clone 5-19 with the recombinant plasmid.

• Parasites, Hosts and Diseases
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• v.34 no.4
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• pp.259-266
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• 1996

The taxonomic validity of morphological group III Accnthamoeba app. is uncertain. In the present study. six type strains of group III Aconthamoeba spry. , A. culbertsoni, A. heniyi, A. pustulosc, A. palestinensis, A. royrebn and A. lenticulnto were subjected for the evaluation or their taxonomic validity by comparison of the isoeneyme patterns by isoelectic focusing on polyacrylamide gels, mitochondrial DNA (Mt DNA) restriction fragment length polymorphism (RFLP) . and small subunit ribosomal DNA (ssu rDNA) PCR-RFLP patterns. The Mt DNA RFLP patterns were heterogeneous between the species. The type strains of A. pclestinensls and A. pustulosc showed almost identical patterns of isoenrymes and rDNA PCR-RFLP with an estimated sequence divergence of 2.6%. The other species showed heterogeneous patterns of isoenxymes and rDNA PCR- RFLP. It is likely that A. pustuLosc is closely related with A. palestinensis and that the former may be regarded as a junior synonym of the latter.

• ALGAE
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• v.20 no.1
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• pp.25-30
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• 2005

To clarify some confusions concerning identification of the Korean Peridinium species, genotypic analysis was performed with their SSU rDNA sequences. PCR was used to amplify the partial SSU rDNA of Peridinium isolates collected from three different Korean waters (Juam, Sang-sa and Togyo Reservoirs). The PCR products were allowed directly to sequence, which revealed each 942 bp of rDNA sequence. Analyses of the rDNA sequences showed that all the Korean isolates had the same genotype (100% sequence homology), and they were nearly identical to a Japanese strain of P. bipes f. occultatum (NIES 364; 99.8% sequence similarity). The sequence-based comparisons could clearly resolve P. bipes f. occultatum isolated from three different Korean waters.

• Journal of Microbiology and Biotechnology
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• v.18 no.5
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• pp.815-820
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• 2008

Two culture-independent methods, namely ribosomal DNA libraries and denaturing gradient gel electrophoresis (DGGE), were adopted to examine the microbial community of a Malaysian light crude oil. In this study, both 16S and 18S rDNAs were PCR-amplified from bulk DNA of crude oil samples, cloned, and sequenced. Analyses of restriction fragment length polymorphism (RFLP) and phylogenetics clustered the 16S and 18S rDNA sequences into seven and six groups, respectively. The ribosomal DNA sequences obtained showed sequence similarity between 90 to 100% to those available in the GenBank database. The closest relatives documented for the 16S rDNAs include member species of Thermoincola and Rhodopseudomonas, whereas the closest fungal relatives include Acremonium, Ceriporiopsis, Xeromyces, Lecythophora, and Candida. Others were affiliated to uncultured bacteria and uncultured ascomycete. The 16S rDNA library demonstrated predomination by a single uncultured bacterial type by >80% relative abundance. The predomination was confirmed by DGGE analysis.