• 제목/요약/키워드: DNA analysis

검색결과 6,599건 처리시간 0.038초

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
    • 대한의생명과학회지
    • /
    • 제26권4호
    • /
    • pp.275-287
    • /
    • 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.

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

  • 김수야;주훈;김재곤;조남표;백병주
    • 대한소아치과학회지
    • /
    • 제23권2호
    • /
    • pp.503-524
    • /
    • 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.

  • PDF

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

  • 기장서;조수연;한명수
    • ALGAE
    • /
    • 제20권1호
    • /
    • pp.25-30
    • /
    • 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
    • /
    • 제8권4호
    • /
    • pp.221-226
    • /
    • 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.

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

  • 송정영;김홍기
    • 한국균학회지
    • /
    • 제30권1호
    • /
    • pp.66-72
    • /
    • 2002
  • Phytophthora capsici 집단의 유전적 특성 분석용 DNA marker를 선발하고자 HindIII로 처리된 P. capsici 95CY3119 균주의 genomic DNA library를 임의로 cloning 시킨 후 southern blot 분석을 실시하여 선발된 clone들의 특성을 조사하였다. Probe로 사용하기 위해 선발된 clone 내에 삽입된 DNA 단편들은 HindIII로 처리된 P. capsici 95CY3119의 genomic DNA와 특이적으로 강하게 반응했다. 조사된 probe 중 PC9은 HindIII로 처리된 국내 P. capsici 균주들의 genomic DNA와 Southern 분석시 많은 밴드를 형성하였으며, 분리포장 단위별 균주들 간에는 물론 동일 포장 분리균주들 간에도 그 차이를 나타냈다. 그 밴드 양상을 기초로 집괴분석을 실시한 결과, 각 균주의 유전적 다양성이 잘 나타났다. Prove PC22는 다른 Phytophthora속과 Pythium속 균주들의 genomic DNA들과의 Southern hybridization에서는 반응을 나타내지 않았지만 특이적으로 P. capsici 균주들과는 다수의 밴드를 형성하였다. 이들 P. capsici 종특이적 DNA probe들은 추후 국내 및 전세계에 분포하는 P. capsici 집단의 유전적 다양성 분석 및 종동정에 유용한 marker로서 활용될 수 있을 것이다.

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

  • 김성광
    • Journal of Yeungnam Medical Science
    • /
    • 제19권1호
    • /
    • pp.1-10
    • /
    • 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.

  • PDF

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

  • 양영렬;허철구
    • KSBB Journal
    • /
    • 제16권6호
    • /
    • pp.538-546
    • /
    • 2001
  • 효모의 세포주기 관련 유전자 발현 통합 데이터를 사용하여 본 연구실에서 개발한 유전자 발현 통합 분석프로그램을 사용하여, 클러스터링 알고리즘의 성능을 비교하고 데이터내에 존재하는 클러스터 개수를 추정하기 위해 FOM 분석을 적용하였으며, 이 분석방법을 통하여 K-means, SOM, Fuzzy c-means 클러스터링 방법의 성능을 서로 비교하였다. 클러스터 개수를 추정한 다음 3가지 클러스터링 방법에 대한 클러스터링 결과 비교, 클러스터의 기능할당 및 모티프 분석을 시도하였다. 본 논문에서 제시하는 분석 방법은 DNA chip 발현 데이터의 일반적인 분석방법을 유전자 발현 패턴의 유사성을 토대로 한 클러스터링 방법에 근간을 두고 있다. 본 논문에서는 클러스터링한 후 각 클러스터의 기능할당 및 모티프 분석에 대한 일반적인 분석방법을 제시하였으며, 본 연구실에서 개발한 유전자 발현분석 통합 프로그램이 효율적으로 사용될 수 있음을 보여주고 있다.

  • PDF

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
    • 한국진공학회:학술대회논문집
    • /
    • 한국진공학회 2013년도 제45회 하계 정기학술대회 초록집
    • /
    • pp.276-276
    • /
    • 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.

  • PDF

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

  • 양영렬;허철구
    • KSBB Journal
    • /
    • 제16권4호
    • /
    • pp.381-388
    • /
    • 2001
  • DNA칩의 유전자 발현 데이터의 통합적 분석을 위하여 매트랩을 기반으로 한 통합분석 프로그램을 구축하였다. 이 프로그램은 유전자 발현 분석을 위해 일반적으로 많이 쓰는 방법인 Hierarchical clustering(HC), K-means, Self-organizing map(SOM), Principal component analysis(PCA)를 지원하며, 이외에 Fuzzy c-means방법과 최근에 발표된 Singular value decomposition(SVD) 분석 방법도 지원하고 있다. 통합분석프로그램의 성능을 알아보기 위하여 효모의 포자형성(sporulation)과 정의 유전자발현 데이터를 사용하였으며, 각 분석 방법에 따른 분석 결과를 제시하였으며, 이 프로그램이 유전자 발현데이타의 통합적인 분석을 위해 효과적으로 사용될 수 있음을 제시하였다.

  • PDF