• Title/Summary/Keyword: DNA analysis

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Validation of Reduced-volume Reaction in the PowerQuant® System for human DNA Quantification

  • Kim, Hyojeong;Cho, Yoonjung;Kim, Jeongyong;Lee, Ja Hyun;Kim, Hyo Sook;Kim, Eungsoo
    • Biomedical Science Letters
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    • v.26 no.4
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    • pp.275-287
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    • 2020
  • Since its introduction in the forensic field, quantitative PCR (qPCR) has played an essential role in DNA analysis. Quality of DNA should be evaluated before short tandem repeat (STR) profiling to obtain reliable results and reduce unnecessary costs. To this end, various human DNA quantification kits have been developed. Among these kits, the PowerQunat® System was designed not only to determine the total amount of human DNA and human male DNA from a forensic evidence item, but also to offer data about degradation of DNA samples. However, a crucial limitation of the PowerQunat® System is its high cost. Therefore, to minimize the cost of DNA quantification, we evaluated kit performance using a reduced volume of reagents (1/2-volume) using DNA samples of varying types and concentrations. Our results demonstrated that the low-volume method has almost comparable performance to the manufacturer's method for human DNA quantification, human male DNA quantification, and DNA degradation index. Furthermore, using a reduced volume of regents, it is possible to run 2 times more reactions per kit. We expect the proposed low-volume method to cut costs in half for laboratories dealing with large numbers of DNA samples.

APPLICATION OF DUAL PARAMETER ANALYSIS IN FLOW CYTOMETRIC DNA MEASUREMENTS OF ORAL SQUAMOUS CELL CARCINOMA (구강편평세포암종의 유세포분석적 DNA측정을 위한 이중 매개변수법의 적용)

  • Kim, Su-Ya;Ju, Hoon;Kim, Jae-Gon;Cho, Nam-Pyo;Baik, Byeong-Ju
    • Journal of the korean academy of Pediatric Dentistry
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    • v.23 no.2
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    • pp.503-524
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    • 1996
  • A series of 31 patients with primary oral squamous cell carcinoma (SCC) who were treated at Chonbuk National University Hospital during the years 1991-1995, were evaluated by dual parameter analysis in flow cytometric DNA measurement, Bryne's malignancy grading system, and the TNM classification. The aims of the present study were to discover that previously undetected aneuploid clones could be detected by dual parameter analysis and to determine the prognostic value of the above parameters. 1. Using dual parameter analysis of cytokeratin and DNA on disintegrated paraffin-embedded samples, aneuploid clones which were undetected by regular single parameter DNA analysis could be found among the cytokeratin-selected cells. DNA aneuploidy from paraffin-embedded samples were 15 cases compared with 10 cases using conventional DNA analysis. 2. The portion of aneuploid tumors showed slightly higher clinical stage and tumor size than the portion of diploid tumors, but the difference was not significant. The portion of DNA aneuploid tumors showed significantly higher mean mitosis and total malignancy scores than the portion of DNA diploid tumors. 3. The majority of the patients presented with clinical stage III and IV lesions showed significantly higher mean total malignancy score as compared to those with clinical stage I and II. 4. Histopathologic mean total malignancy score of the 31 cases was 12.7. Among the histologic parameters, mean mitosis score was correlated to the status of DNA ploidy and total malignancy score were correlated to the DNA ploidy and clinical staging.

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Analysis of Small-Subunit rDNA Sequences Obtained from Korean Peridinium bipes f. occultatum (Dinophyceae) (한국산 와편모조류 Peridinium bipes f. occultatum의 Small-Subunit Ribosomal DNA(SSU rDNA) 염기서열 분석)

  • Ki, Jang-Seu;Cho, Soo-Yeon;Han, Myung-Soo
    • 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.

Nanoparticle-based Detection Technology for DNA Analysis

  • Park, Hyun-Gyu
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.8 no.4
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    • pp.221-226
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    • 2003
  • With the current rapid development of nanotechnology and synthesis technology for designed oligonucleotides or oligonucleotide-modified nanoparticle conjugates, the combined strategies have become one of the most valuable methods in detection technology for DNA analysis. Using the uniquely recognizable interactions of pre-designed DNA molecules in assembling nanoparticles, various novel approaches have been recently developed towards detecting specific DNA sequences. Here we describe the key fundamentals and issues of this promising strategies ranging from the initial findings of rationally designed DNA-based assembly of nanoparticles to the extended chip-based detection system. Some limitations of these new strategies and possible approaches will be also discussed for the practical application in the area of DNA microarray detection.

Development of Repetitive DNA Probes for Genetic Analysis of Phytophthora capsici (Phytophthora capsici의 유전적 특성 분석을 위한 Repetitive DNA Probe의 개발)

  • Song, Jeong-Young;Kim, Hong-Gi
    • The Korean Journal of Mycology
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    • v.30 no.1
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    • pp.66-72
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    • 2002
  • To develop DNA markers for analysis of genetic characteristics of Phytophthora capsici population, randomly selected clones from HindIII-digested genomic DNA library of P. capsici 95CY3119 were surveyed by hybridizing to Southern blots of HindIII-digested total genomic DNA of P. capsici. Probe DNAs inserted into selected individual clones strongly hybridized with HindIII digests of P. capsici. Among probes examined, PC9 revealed the repetitive and highly polymorphic bands to HindIII digests of inter-and intra-field P. capsici isolates. Genetic diversity of individual isolates was also clearly revealed in cluster analysis based on its band patterns. The other probe, PC22, was hybridized only to DNA from P. capsici and this was highly repetitive. However, there was no response to other Phytophthora species and Pythium sp. These DNA probes could be used as very useful markers in analysing genetic diversity and identification for P. capsici population throughout the world.

Microbial Genome Analysis and Application to Clinical Bateriology (미생물의 유전자(Genome) 해석과 임상세균학에 이용)

  • Kim, Sung-Kwang
    • Journal of Yeungnam Medical Science
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    • v.19 no.1
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    • pp.1-10
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    • 2002
  • With the establishment of rapid sequence analysis of 16S rRNA and the recognition of its potential to determine the phylogenetic position of any prokaryotic organism, the role of 16S rRNA similarities in the present species definition in bacteriology need to be clarified. Comparative studies clearly reveal the limitations of the sequence analysis of this conserved gene and gene product in the determination of relationship at the pathogenic strain level for which DNA-DNA reassociation experiments still constitute the superior method. Since today the primary structure of 16S rRNA is easier to determine than hybridization between DNA strands, the strength of the sequence analysis is to recognize the level at which DNA pairing studies need to be performed, which certainly applies to similarities of 97% and higher.

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An integrated DNA barcode assay microdevice for rapid, highly sensitive and multiplex pathogen detection at the single-cell level

  • Jung, Jae Hwan;Cho, Min Kyung;Chung, So Yi;Seo, Tae Seok
    • Proceedings of the Korean Vacuum Society Conference
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    • 2013.08a
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    • pp.276-276
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    • 2013
  • Here we report an integrated microdevice consisting of an efficient passive mixer, a magnetic separation chamber, and a capillary electrophoretic microchannel in which DNA barcode assay, target pathogen separation, and barcode DNA capillary electrophoretic analysis were performed sequentially within 30 min for multiplex pathogen detection at the single-cell level. The intestine-shaped serpentine 3D micromixer provides a high mixing rate to generate magnetic particle-pathogenic bacteria-DNA barcode labelled AuNP complexes quantitatively. After magnetic separation and purification of those complexes, the barcode DNA strands were released and analyzed by the microfluidic capillary electrophoresis within 5 min. The size of the barcode DNA strand was controlled depending on the target bacteria (Staphylococcus aureus, Escherichia coli O157:H7, and Salmonella typhimurium), and the different elution time of the barcode DNA peak in the electropherogram allows us to recognize the target pathogen with ease in the monoplex as well as in the multiplex analysis. In addition, the quantity of the DNA barcode strand (~104) per AuNP is enough to be observed in the laser-induced confocal fluorescence detector, thereby making single-cell analysis possible. This novel integrated microdevice enables us to perform rapid, sensitive, and multiplex pathogen detection with sample-in-answer-out capability to be applied for biosafety testing, environmental screening, and clinical trials.

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Analysis of Combined Yeast Cell Cycle Data by Using the Integrated Analysis Program for DNA chip (DNA chip 통합분석 프로그램을 이용한 효모의 세포주기 유전자 발현 통합 데이터의 분석)

  • 양영렬;허철구
    • KSBB Journal
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    • v.16 no.6
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    • pp.538-546
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    • 2001
  • An integrated data analysis program for DNA chip containing normalization, FDM analysis, various kinds of clustering methods, PCA, and SVD was applied to analyze combined yeast cell cycle data. This paper includes both comparisons of some clustering algorithms such as K-means, SOM and furry c-means and their results. For further analysis, clustering results from the integrated analysis program was used for function assignments to each cluster and for motif analysis. These results show an integrated analysis view on DNA chip data.

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Program Development of Integrated Expression Profile Analysis System for DNA Chip Data Analysis (DNA칩 데이터 분석을 위한 유전자발연 통합분석 프로그램의 개발)

  • 양영렬;허철구
    • KSBB Journal
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    • v.16 no.4
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    • pp.381-388
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    • 2001
  • A program for integrated gene expression profile analysis such as hierarchical clustering, K-means, fuzzy c-means, self-organizing map(SOM), principal component analysis(PCA), and singular value decomposition(SVD) was made for DNA chip data anlysis by using Matlab. It also contained the normalization method of gene expression input data. The integrated data anlysis program could be effectively used in DNA chip data analysis and help researchers to get more comprehensive analysis view on gene expression data of their own.

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