• Title/Summary/Keyword: DNA-Based Methods

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Evaluation of DNA Extraction Methods from Low Copy Number (LCN) DNA Samples for Forensic DNA Typing

  • Eom, Yong-Bin
    • 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.

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Application of Methodology for Microbial Community Analysis to Gas-Phase Biofilters (폐가스 처리용 바이오필터에 미생물 군집 분석 기법의 적용)

  • Lee, Eun-Hee;Park, Hyunjung;Jo, Yun-Seong;Ryu, Hee Wook;Cho, Kyung-Suk
    • Korean Chemical Engineering Research
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    • v.48 no.2
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    • pp.147-156
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    • 2010
  • There are four key factors for gas-phase biofilters; biocatalysts(microorganisms), packing materials, design/operating techniques, and diagnosis/management techniques. Biofilter performance is significantly affected by microbial community structures as well as loading conditions. The microbial studies on biofilters are mostly performed on basis of culture-dependent methods. Recently, advanced methods have been proposed to characterize the microbial community structure in environmental samples. In this study, the physiological, biochemical and molecular methods for profiling microbial communities are reviewed, and their applicability to biofilters is discussed. Community-level physiological profile is based on the utilization capability of carbon substrate by heterotrophic community in environmental samples. Phospholipid fatty acid analysis method is based on the variability of fatty acids present in cell membranes of different microorganisms. Molecular methods using DNA directly extracted from environmental samples can be divided into "partial community DNA analysis" and "whole community DNA analysis" approaches. The former approaches consist in the analysis of PCR-amplified sequence, the genes of ribosomal operon are the most commonly used sequences. These methods include PCR fragment cloning and genetic fingerprinting such as denaturing gradient gel electrophoresis, terminal-restriction fragment length polymorphism, ribosomal intergenic spacer analysis, and random amplified polymorphic DNA. The whole community DNA analysis methods are total genomic cross-DNA hybridization, thermal denaturation and reassociation of whole extracted DNA and extracted whole DNA fractionation using density gradient.

Advantages of the single nucleotide polymorphism-based noninvasive prenatal test

  • Kim, Kunwoo
    • Journal of Genetic Medicine
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    • v.12 no.2
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    • pp.66-71
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    • 2015
  • Down syndrome screening with cell-free DNA (cfDNA) in the maternal plasma has recently received much attention in the prenatal diagnostic field. Indeed, a large amount of evidence has already accumulated to show that screening tests with cfDNA are more sensitive and specific than conventional maternal serum and/or ultrasound screening. Globally, more than 1,000,000 of these noninvasive prenatal tests (NIPTs) have been performed to date. There are several different methods for NIPTs that are currently commercially available, including shotgun massively parallel sequencing, targeted massively parallel sequencing, and single nucleotide polymorphism (SNP)-based methods. All of these methods have their own advantages and disadvantages. In this review, I will focus specifically on the SNP-based NIPT.

Evaluation of Methods to Analyze SNP-based Association Studies in a DNA-Pooling Experiment with Preferential Amplification

  • Ahn, Chul;Lee, Kyu-Sang
    • Proceedings of the Korean Society for Bioinformatics Conference
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    • 2005.09a
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    • pp.395-398
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    • 2005
  • Genetic association case-control studies using DNA pools are efficient ways of detecting association between a marker allele and disease status. DNA pooling is an efficient screening method for locating susceptibility genes associated with the disease. However, DNA pooling is efficient only when allele frequency estimation is done precisely and accurately. Through the evaluation of empirical type I errors and empirical powers by simulation, we will evaluate the methods that correct for preferential amplification of nucleotides when estimating the allele frequency of single-nucleotide polymorphisms.

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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.

Comparison of Non-amplified and Amplified DNA Preparation Methods for Array-comparative Gnomic Hybridization Analysis

  • Joo, Hong-Jin;Jung, Seung-Hyun;Yim, Seon-Hee;Kim, Tae-Min;Xu, Hai-Dong;Shin, Seung-Hun;Kim, Mi-Young;Kang, Hyun-Mi;Chung, Yeun-Jun
    • Molecular & Cellular Toxicology
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    • v.4 no.3
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    • pp.246-252
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    • 2008
  • Tumor tissue is usually contaminated by normal tissue components, which reduces the sensitivity of analysis for exploring genetic alterations. Although microdissection has been adopted to minimize the contamination of tumor DNA with normal cell components, there is a concern over the amount of microdissected DNA not enough to be applied to array-CGH reaction. To amplify the extracted DNA, several whole genome amplification (WGA) methods have been developed, but objective comparison of the array-CGH outputs using different types of WGA methods is still scarce. In this study, we compared the performance of non-amplified microdissected DNA and DNA amplified in 2 WGA methods such as degenerative oligonucleotide primed (DOP)-PCR, and multiple strand displacement amplification (MDA) using Phi 29 DNA polymerase. Genomic DNA was also used to make a comparison. We applied those 4 DNAs to whole genome BAC array to compare the false positive detection rate (FPDR) and sensitivity in detecting copy number alterations under the same hybridization condition. As a result microdissected DNA method showed the lowest FPDR and the highest sensitivity. Among WGA methods, DOP-PCR amplified DNA showed better sensitivity but similar FPDR to MDA-amplified method. These results demonstrate the advantage and applicability of microdissection for array-CGH analysis, and provide useful information for choosing amplification methods to study copy number alterations, especially based on precancerous and microscopically invaded lesions.

Current methodologies in construction of plant-pollinator network with emphasize on the application of DNA metabarcoding approach

  • Namin, Saeed Mohamadzade;Son, Minwoong;Jung, Chuleui
    • Journal of Ecology and Environment
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    • v.46 no.2
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    • pp.126-135
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    • 2022
  • Background: Pollinators are important ecological elements due to their role in the maintenance of ecosystem health, wild plant reproduction, crop production and food security. The pollinator-plant interaction supports the preservation of plant and animal populations and it also improves the yield in pollination dependent crops. Having knowledge about the plant-pollinator interaction is necessary for development of pesticide risk assessment of pollinators and conservation of endangering species. Results: Traditional methods to discover the relatedness of insects and plants are based on tracing the visiting pollinators by field observations as well as palynology. These methods are time-consuming and needs expert taxonomists to identify different groups of pollinators such as insects or identify flowering plants through palynology. With pace of technology, using molecular methods become popular in identification and classification of organisms. DNA metabarcoding, which is the combination of DNA barcoding and high throughput sequencing, can be applied as an alternative method in identification of mixed origin environmental samples such as pollen loads attached to the body of insects and has been used in DNA-based discovery of plant-pollinator relationship. Conclusions: DNA metabarcoding is practical for plant-pollinator studies, however, lack of reference sequence in online databases, taxonomic resolution, universality of primers are the most crucial limitations. Using multiple molecular markers is preferable due to the limitations of developed universal primers, which improves taxa richness and taxonomic resolution of the studied community.

DNA Coding Method for Time Series Prediction (시계열 예측을 위한 DNA 코딩 방법)

  • 이기열;선상준;이동욱;심귀보
    • 제어로봇시스템학회:학술대회논문집
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    • 2000.10a
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    • pp.280-280
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    • 2000
  • In this paper, we propose a method of constructing equation using bio-inspired emergent and evolutionary concepts. This method is algorithm that is based on the characteristics of the biological DNA and growth of plants. Here is. we propose a constructing method to make a DNA coding method for production rule of L-system. L-system is based on so-called the parallel rewriting mechanism. The DNA coding method has no limitation in expressing the production rule of L-system. Evolutionary algorithms motivated by Darwinian natural selection are population based searching methods and the high performance of which is highly dependent on the representation of solution space. In order to verify the effectiveness of our scheme, we apply it to one step ahead prediction of Mackey-Glass time series.

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A DNA Coding-Based Interacting Multiple Model Method for Tracking a Maneuvering Target (기동 표적 추적을 위한 DNA 코딩 기반 상호작용 다중모델 기법)

  • Lee, Bum-Jik;Joo, Young-Hoon;Park, Jin-Bae
    • Proceedings of the KIEE Conference
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    • 2002.11c
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    • pp.87-91
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    • 2002
  • The problem of maneuvering target tracking has been studied in the field of the state estimation over decades. The Kalman filter has been widely used to estimate the state of the target, but in the presence of a maneuver, its performance may be seriously degraded. In this paper, to solve this problem and track a maneuvering target effectively, a DNA coding-based interacting multiple model (DNA coding-based IMM) method is proposed. The proposed method can overcome the mathematical limits of conventional methods by using the fuzzy logic based on DNA coding method. The tracking performance of the proposed method is compared with those of the adaptive IMM algorithm and the GA-based IMM method in computer simulations.

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DNA Coding-Based Intelligent Kalman Filter for Tracking a Maneuvering Target (기동표적 추적을 위한 DNA 코딩 기반 지능형 칼만 필터)

  • 이범직;주영훈;박진배
    • Proceedings of the Korean Institute of Intelligent Systems Conference
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    • 2002.12a
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    • pp.118-121
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    • 2002
  • The problem of maneuvering target tracking has been studied in the field of the state estimation over decades. The Kalman filter has been widely used to estimate the state of the target, but in the presence of a maneuver, its performance may be seliously degraded. In this paper, to solve this problem and track a maneuvering target effectively, DNA coding-based intelligent Kalman filter (DNA coding-based IKF) is proposed. The proposed method can overcome the mathematical limits of conventional methods and can effectively track a maneuvering target with only one filter by using the fuzzy logic based on DNA coding method. The tracking performance of the proposed method is compared with those of the adaptive interacting multiple model (AIMM) method and the GA-based IKF in computer simulations.