• Title/Summary/Keyword: DNA barcode analysis

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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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Identification of bird species and their prey using DNA barcode on feces from Korean traditional village groves and forests (maeulsoop)

  • Joo, Sungbae;Park, Sangkyu
    • Animal cells and systems
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    • v.16 no.6
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    • pp.488-497
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    • 2012
  • A DNA barcode based on 648 bp of cytochrome c oxidase I (COI) gene aims to build species-specific libraries for animal groups. However, it is hard to recover full-length (648 bp) barcode gene from environmental fecal samples due to DNA degradation. In this study, we designed a new primer set (K_Bird), which amplifies a 226 bp fragment targeted an inner position of full-length COI barcode based on 102 species of Korean birds to improve amplification success, and we attempted to identify bird species from 39 avian fecal samples collected during 4 months from Jinan, South Korea. Simultaneously, we conducted a dietary analysis using a universal DNA mini-barcode (Uni_Minibar) from same fecal samples. In silico analysis on newly designed mini-barcode represented that genetic distances were 0.5% in species and 9.1% in genera. Intraspecific variations of 149 species out of 174 species (86%) between Korea and North America were within the threshold (5.3% threshold in this study). From environmental fecal samples collected in Jinan, we identified seven avian species, which have high similarity (99-100%) with registered COI sequences in GenBank. Eight kinds of prey species, such as moth, spider, fly, and dragonfly, were identified in dietary analysis. We suppose that our strategy applying mini-barcode for environmental fecal samples, might be a useful and convenient tool for species identification and dietary analysis for birds.

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.

Error Correcting using the Check digit on Barcode, and the present and future of Barcode (바코드에 있어서 체크숫자를 이용한 오류수정과 바코드의 현재와 미래)

  • Kim, Hwa-Joon
    • Journal for History of Mathematics
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    • v.21 no.2
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    • pp.103-118
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    • 2008
  • Barcode technology is becoming an essential tool for every companies, and this makes help us to gain time, analysis of goods, an inventory control, a prevention of burglar and so on. In this paper, we have treated about the history of barcode, its error correcting using the check digit, and the present and future of barcode. We wish roles of mathematician on new barcode system which it bring on an economical efficiency and stability.

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Phylogenetic analysis of 14 Korean Araliaceae species using chloroplast DNA barcode analysis (엽록체 DNA 바코드 분석을 통한 한국산 두릅나무과 식물 14종의 유연관계 분석)

  • Hwang, Hwan Su;Choi, Yong Eui
    • Journal of Plant Biotechnology
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    • v.43 no.1
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    • pp.82-90
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    • 2016
  • Most Araliaceae plant species distributed in Korea are economically important because of their high medicinal values. This study was conducted to develop barcode markers from sequence analysis of chloroplast DNA in 14 taxa of Araliaceae species grown in South Korea. Sequencing of seven chloroplast DNA regions was performed to establish the DNA barcode markers, as suggested by the Consortium for the Barcode of Life (CBOL). From the sequence analysis of chloroplast DNA, we identified specific sequences and nucleotides that allowed us to discriminate among each other 14 Korean Araliaceae species. The sequence in the region of psbA-trnH revealed the most frequent DNA indels and substitutions of all 7 regions studied. This psbA-trnH marker alone can discriminate among all 14 species. There are no differences between Korean and Chinese Panax ginseng in all seven sequenced chloroplast DNA regions. A phylogenetic tree constructed using the seven chloroplast DNA regions revealed that Tetrapanax papyriferus should be classified as an independent clade. The Aralia and Panax genera showed a close phylogenetic relationship. Five species in the Eleutherococcus genus were more closely related to Kalopanax septemlobus than to any Panax species.

Identification of Marker Nucleotides for the Molecular Authentication of Arisaematis Rhizoma Based on the DNA Barcode Sequences (천남성(天南星) 유전자 감별을 위한 DNA 바코드 분석 및 Marker Nucleotide 발굴)

  • Kim, Wook Jin;Lee, Young Mi;Ji, Yunui;Kang, Young Min;Choi, Goya;Kim, Ho Kyoung;Moon, Byeong Cheol
    • The Korea Journal of Herbology
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    • v.29 no.6
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    • pp.35-43
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    • 2014
  • Objectives : Official Arisaematis Rhizoma is described only three species, Arisaema amurnse, Arisaema erubescens, and Arisaema heterophyllum, in national Pharmacopoeia. However, other Arisaema species, Arisaema ringens, Arisaema takesimense and Arisaema serratum, also have been distributed as an inauthentic Arisaematis Rhizoma in the herbal market. To develop a reliable molecular authentication method for Arisaematis Rhizoma in species level, we analyzed DNA barcode regions using six Arisaema species. Methods : Thirty-eight samples of six Arisaema plants species (A. amurense, A. amurense f. serratum, A. heterophyllum, A. takesimense, and A. serratum) were collected from different habitate and nucleotide sequences of DNA barcode regions (rDNA-ITS, matK, and rbcL gene) were analyzed after PCR amplification. The species-specific sequences and phylogenetic relations were estimated using entire sequences of three DNA barcodes based on the analysis of ClastalW and UPGMA, respectively. Results : The comparative analysis of DNA barcode sequences were revealed inter-species specific nucleotides to distinguish the medicinal plant of Arisaema Rhizoma in species levels excluding between A. amurense and its subspecies (A. amurense f. serratum) and A. takesimense and A. serratum, respectively. However, we obtained sequence differences enough to discriminate authentic and inauthentic Arisaematis Rhizoma. Therefore, we suggest that these SNP type molecular genetic markers were an reliable method avaliable to identify official herbal medicines. Conclusions : These marker nucleotides could be useful to identify the official herbal medicines by providing definitive information that can identify original medicinal plant and distinguish from inauthentic adulterants and substitutes.

Development of Molecular Markers for the authentication of Zanthoxyli Pericarpium by the analysis of rDNA-ITS DNA barcode regions (rDNA-ITS DNA 바코드 부위 분석을 통한 산초(山椒) 기원종 감별용 유전자 마커 개발)

  • Kim, Wook Jin;Ji, Yunui;Lee, Young Mi;Kang, Young Min;Choi, Goya;Moon, Byeong Cheol
    • The Korea Journal of Herbology
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    • v.30 no.3
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    • pp.41-47
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    • 2015
  • Objectives : Due to the morphological similarity of the pericarp and description of multi-species in National Pharmacopoeia of Korea and China, the Zanthoxylum Pericarpium is difficult to authenticate adulterant in species levels. Therefore, we introduced the sequence analysis of DNA barcode and identification of single nucleotide polymorphism(SNP) to establish a reliable tool for the distinction of Zanthoxylum Pericarpium from its adulterants. Methods : To analyze DNA barcode region, genomic DNA was extracted from twenty-four specimens of authentic Zanthoxylum species and inauthentic adulterant and the individual internal transcribed spacer regions (rDNA-ITS and ITS2) of nuclear ribosomal RNA gene were amplified using ITS1, ITS2-S2F, and ITS4 primer. For identification of species-specific sequences, a comparative analysis was performed using entire DNA barcode sequences. Results : In comparison of four Zanthoxylum ITS2 sequences, we identified 16, 4, 6, and 4 distinct species-specific nucleotides enough to distinguish Z. schinifolium, Z. bungeanum, Z. piperitum, and Z. simulans, respectively. The sequence differences were available genetic marker to discriminate four species. Futhermore, phylogenetic relationship revealed a clear classification between different Zanthoxylum species showing 4 different clusters. These results indicated that comparative analysis of ITS2 DNA barcode was an useful genetic marker to authenticate Zanthoxylum Pericarpium in species levels. Conclusions : The marker nucleotides, enough to distinguish Z. schinifolium, Z. piperitum, Z. bungeanum, and Z. simulans, were obtained at 30 SNP marker nucleotides from ITS2 sequences. These differences could be used to authenticate official Zanthoxylum Pericarpium from its adulterants as well as discriminating each four species.

Molecular Authentication of Pinelliae Tuber from its adulterants by the analysis of DNA barcodes, matK and rbcL genes (matK와 rbcL DNA 바코드 분석을 통한 반하(半夏) 및 반하(半夏) 유사 한약재 유전자 감별)

  • Lee, Young Mi;Moon, Byeong Cheol;Ji, Yunui;Kim, Wook Jin;Kim, Ho Kyoung
    • The Korea Journal of Herbology
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    • v.28 no.6
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    • pp.53-58
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    • 2013
  • Objectives : Pinelliae Tuber has been used as a typical unauthentic herbal medicines. Due to the morphological similarity between Pinelliae Tuber and adulterants, the correct authentication is very difficult. Therefore, we introduced DNA barcode to establish a powerful tool for the authentication of Pinelliae Tuner from adulterants. Methods : To obtain DNA barcode regions, genomic DNA was extracted from nineteen specimens of Pinellia ternata, Pinellia pedatisecta, Pinellia tripartita, and Typhonium flagelliforme, and matK and rbcL genes were amplified. For identification of species specific sequences and analysis phylogenetic relationship, a comparative analysis were performed by the ClastalW and UPGMA based on entire sequences of matK and rbcL genes, respectively. Results : In comparison of two DNA barcode sequences, we elucidated the phylogenetic relationship showing distinct four groups depending on species and identified 40 and 20 species specific nucleotides enough to distinguish each species from matK and rbcL gene, respectively. The sequence differences at the corresponding positions were avaliable genetic marker nulceotides to discriminate the correct species among analyzed four species. These results indicated that phylogentic and comparative analysis of matK and rbcL genes are useful genetic markers to authenticate Pinelliae Tubers. Conclusions : The marker nucleotides enough to distinguish P. ternata, P. tripatrita, P. peditisecta, and T. flagelliform, were observed at 40 positions in matK gene and 20 positions in rbcL gene sequence, respectively. These differences can be used to authenticate Pinelliae Tuber from adulterants as well as discriminate each four species.

Molecular Authentication and Phylogenetic Analysis of Plant Species for Breeae and Cirsii Herba based on DNA barcodes (DNA 바코드 분석을 통한 소계(小薊) 및 대계(大薊) 기원식물 감별과 종간 유연관계 분석)

  • Moon, Byeong Cheol;Lee, Young Mi;Ji, Yunui;Choi, Goya;Chun, Jin Mi;Kim, Ho Kyoung
    • The Korea Journal of Herbology
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    • v.28 no.3
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    • pp.75-84
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    • 2013
  • Objectives : The origin of Breeae Herba (So-gye) and Cirsii Herba (Dae-gye) is differently prescribed in Korean and Chinese modern pharmacopoeia. Since the similar morphological characteristics and chaotic plant names, moreover, the aerial part of Carduus crispus have been used as the Cirsii Herba. To develop a reliable method for correct identification of these herbal medicines and to evaluate the genetic relationship of these closely related plant species, we analyzed sequences of DNA barcode regions. Methods : Thirty-one samples of 6 medicinal plants (B. segeta, B. setosa, C. japonicum var. maackii, C. setidens, C. chanroenicum, and C. crispus) were collected from different habitate and nucleotide sequences of DNA barcode regions (rDNA-ITS, matK, and rbcL) were analyzed after amplification using appropriate primers reported in previous studies. The nucleotides of species-specific authentic marker and phylogenetic relations were estimated based on the entire sequences of DNA barcodes by the analysis of ClastalW and UPGMA, respectively. Results : In comparative analysis of DNA barcode sequences, we obtained specific nucleotides to discriminate the medicinal plant of Breeae/Cirsii Herba in species level and evaluated the phylogenetic relationship of these species. Futhermore, we identified distinct marker nucleotides enough to authenticate respective species. These sequence differences at corresponding positions were avaliable genetic markers to determine the botanical origins of Breeae Herbal as well as Cirsii Herba. Conclusions : These marker nucleotides would be useful to identify the official herbal medicines by providing of definitive information that can identify each plant species and distinguish from unauthentic adulterants and substitutes.

The First Record of the Marphysa victori (Polychaeta, Eunicida, Eunicidae) from Korea, with DNA Barcode Data

  • Kim, Hana;Kim, Keun-Yong;Phoo, War War;Kim, Chang-Hoon
    • Animal Systematics, Evolution and Diversity
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    • v.37 no.1
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    • pp.1-8
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    • 2021
  • A eunicid polychaete, Marphysa victori Lavesque, Daffe, Bonifácio & Hutchings, 2017 is described for the first time from the intertidal zones of the Korean coasts. It is characterized by having three types of pectinate chaetae (INS, isodont-narrow-slender; AWS, anodont-wide-slender; and AWT, anodont-wide-thick), appearance of pectinate chaetae from chaetiger 2, the chaetae consisted of pectinate and compound spinigers, and pygidium with one pair of pygidial cirri. In genetic analysis based on cytochrome c oxidase subunit I (COI), intra-specific genetic distance between the specimens of M. victori from its type locality, France and Korea are in the range of 0.000-0.013. This paper includes the morphological description and photographs of M. victori new to Korean fauna, with partial sequences of the mitochondrial COI as DNA barcode data on this species.