• Title/Summary/Keyword: large dna

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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.

A Proteomic Approach to Study msDNA Function in Escherichia coli

  • Jeong, Mi-Ae;Lim, Dongbin
    • Journal of Microbiology
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    • v.42 no.3
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    • pp.200-204
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    • 2004
  • Retron is a prokaryotic genetic element that produces multicopy single-stranded DNA covalently linked to RNA (msDNA) by a reverse transcriptase. It was found that cells producing a large amount of msDNA, rather than those that did not, showed a higher rate of mutation. In order to understand the molecular mechanism connecting msDNA production to the high mutation rate the protein patterns were compared by two dimensional gel electrophoresis. Ten proteins were found to be differentially expressed at levels more than three fold greater in cells with than without msDNA, nine of which were identified by MALDI TOF MS. Eight of the nine identified proteins were repressed in msDNA-producing cells and, surprisingly, most were proteins functioning in the dissimilation of various carbon sources. One protein was induced four fold greater in the msDNA producing cells and was identified as a 30S ribosomal protein S2 involved in the regulation of translation. The molecular mechanism underlying the elevated mutation in msDNA-producing cell still remains elusive.

An assessment of the taxonomic reliability of DNA barcode sequences in publicly available databases

  • Jin, Soyeong;Kim, Kwang Young;Kim, Min-Seok;Park, Chungoo
    • ALGAE
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    • v.35 no.3
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    • pp.293-301
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    • 2020
  • The applications of DNA barcoding have a wide range of uses, such as in taxonomic studies to help elucidate cryptic species and phylogenetic relationships and analyzing environmental samples for biodiversity monitoring and conservation assessments of species. After obtaining the DNA barcode sequences, sequence similarity-based homology analysis is commonly used. This means that the obtained barcode sequences are compared to the DNA barcode reference databases. This bioinformatic analysis necessarily implies that the overall quantity and quality of the reference databases must be stringently monitored to not have an adverse impact on the accuracy of species identification. With the development of next-generation sequencing techniques, a noticeably large number of DNA barcode sequences have been produced and are stored in online databases, but their degree of validity, accuracy, and reliability have not been extensively investigated. In this study, we investigated the extent to which the amount and types of erroneous barcode sequences were deposited in publicly accessible databases. Over 4.1 million sequences were investigated in three largescale DNA barcode databases (NCBI GenBank, Barcode of Life Data System [BOLD], and Protist Ribosomal Reference database [PR2]) for four major DNA barcodes (cytochrome c oxidase subunit 1 [COI], internal transcribed spacer [ITS], ribulose bisphosphate carboxylase large chain [rbcL], and 18S ribosomal RNA [18S rRNA]); approximately 2% of erroneous barcode sequences were found and their taxonomic distributions were uneven. Consequently, our present findings provide compelling evidence of data quality problems along with insufficient and unreliable annotation of taxonomic data in DNA barcode databases. Therefore, we suggest that if ambiguous taxa are presented during barcoding analysis, further validation with other DNA barcode loci or morphological characters should be mandated.

Application of Breed-specific DNA Markers for the use of Identifying Major Pure Pig Breeds Maintained in Korea (대한민국내 주요 돼지 품종의 순종 식별을 위한 품종특이 DNA marker의 활용)

  • Seo, B.Y.;Kim, J.H.;Park, E.W.;Lim, H.T.;Cho, I.C.;Kim, B.W.;Oh, S.J.;Cheong, I.C.;Lee, J.G.;Jeon, J.T.
    • Journal of Animal Science and Technology
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    • v.46 no.5
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    • pp.735-742
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    • 2004
  • This study was conducted for the identification of pure Landrace, Large White and Duroc breeds which are mainly maintained in Korea using DNA markers. We used known KIT and MC1R mutations, which were related coat color in pigs, and pig mitochondrial DNA variations. The KIT mutation was used to distinguish white and colored animals. Duroc breed could be discriminated from other colored breeds using the MC1R mutation N121D. Discriminating Landrace and Large White was possible using the l l-bp duplication of D-Ioop region and alternative initiation codon of ND2. In conclusion, identification of Landrace, Large White and Duroc breeds was might be possible using the procedure designed in this study.

A Space Efficient Indexing Technique for DNA Sequences (공간 효율적인 DNA 시퀀스 인덱싱 방안)

  • Song, Hye-Ju;Park, Young-Ho;Loh, Woong-Kee
    • Journal of KIISE:Databases
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    • v.36 no.6
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    • pp.455-465
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    • 2009
  • Suffix trees are widely used in similar sequence matching for DNA. They have several problems such as time consuming, large space usages of disks and memories and data skew, since DNA sequences are very large and do not fit in the main memory. Thus, in the paper, we present a space efficient indexing method called SENoM, allowing us to build trees without merging phases for the partitioned sub trees. The proposed method is constructed in two phases. In the first phase, we partition the suffixes of the input string based on a common variable-length prefix till the number of suffixes is smaller than a threshold. In the second phase, we construct a sub tree based on the disk using the suffix sets, and then write it to the disk. The proposed method, SENoM eliminates complex merging phases. We show experimentally that proposed method is effective as bellows. SENoM reduces the disk usage less than 35% and reduces the memory usage less than 20% compared with TRELLIS algorithm. SENoM is available to query efficiently using the prefix tree even when the length of query sequence is large.

A Versatile Method for DNA Sequencing of Unpurified PCR Products using an Automated DNA Sequencer and Tailed or Nested Primer Labeled with Near-infrared Dye: A Case Study on the Harmful Dinoflagellate Alexandrium

  • Ki Jang-Seu;Han Myung-Soo
    • Fisheries and Aquatic Sciences
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    • v.9 no.2
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    • pp.70-74
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    • 2006
  • DNA sequence-based typing is considered a robust tool for the discrimination of dinoflagellate species because of the availability of extensive rDNA sequences. Here, we present a rapid, cost-effective DNA-sequencing technique for various PCR products. This sequencing strategy relies on 'nested' or 'tailed' primer labeled with near-infrared dye, and uses a minimal volume of unpurified PCR product (ca. $5{\mu}L$) as the DNA template for sequencing reactions. Reliable and accurate base identification was obtained for several hundred PCR fragments of rRNA genes. This quick, inexpensive technique is widely applicable to sequence-based typing in clinical applications, as well as to large-scale DNA sequencing of the same genomic regions from related species for studies of molecular evolution.

DNA Metallization for Nanoelectronics (DNA 기반 금속 나노 와이어의 제작기술)

  • Han, Gyeongyeop;Lee, Jungkyu K.
    • Applied Chemistry for Engineering
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    • v.29 no.3
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    • pp.253-257
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    • 2018
  • DNA metallization has been emerged as a candidate for fabricating nanocircuits because of its simple process over a large area on a surface. With unique properties, DNA can be an excellent template to achieve molecular electronics. Thus, we introduced the preparation and properties of DNA metallization, and also suggested future directions in this review.

On the continuum formulation for modeling DNA loop formation

  • Teng, Hailong;Lee, Chung-Hao;Chen, Jiun-Shyan
    • Interaction and multiscale mechanics
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    • v.4 no.3
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    • pp.219-237
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    • 2011
  • Recent advances in scientific computing enable the full atomistic simulation of DNA molecules. However, there exists length and time scale limitations in molecular dynamics (MD) simulation for large DNA molecules. In this work, a two-level homogenization of DNA molecules is proposed. A wavelet projection method is first introduced to form a coarse-grained DNA molecule represented with superatoms. The coarsened MD model offers a simplified molecular structure for the continuum description of DNA molecules. The coarsened DNA molecular structure is then homogenized into a three-dimensional beam with embedded molecular properties. The methods to determine the elasticity constants in the continuum model are also presented. The proposed continuum model is adopted for the study of mechanical behavior of DNA loop.

DNA Sequence Visualization with k-convex Hull (k-convex hull을 이용한 DNA 염기 배열의 가시화)

  • Kim, Min Ah;Lee, Eun Jeong;Cho, Hwan Gyu
    • Journal of the Korea Computer Graphics Society
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    • v.2 no.2
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    • pp.61-68
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    • 1996
  • In this paper we propose a new visualization technique to characterize qualitative information of a large DNA sequence. While a long DNA sequence has huge information, it is not easy to obtain genetic information from the DNA sequence. We transform DNA sequences into a polygon to compute their homology in image domain rather than text domain. Our program visualizes DNA sequences with colored random walk plots and simplify them k-convex hulls. A random walk plot represents DNA sequence as a curve in a plane. A k-convex hull simplifies a random work plot by removing some parts of its insignificant information. This technique gives a biologist an insight to detect and classify DNA sequences with easy. Experiments with real genome data proves our approach gives a good visual forms for long DNA sequences for homology analysis.

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Principle of Emulsion PCR and Its Applications in Biotechnology

  • Chai, Changhoon
    • Journal of Animal Reproduction and Biotechnology
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    • v.34 no.4
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    • pp.259-266
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    • 2019
  • Emulsion polymerase chain reaction (PCR) is performed on compartmentalized DNA, allowing a large number of PCR reactions to be carried out in parallel. Emulsion PCR has unique advantages in DNA amplification. It can be applied in many molecular biological assays, especially those requiring highly sensitive and specific DNA amplification. This review discusses the principle of emulsion PCR and its applications in biotechnology. Related technologies are also discussed.