• Title/Summary/Keyword: Illumina HiSeq platform

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Comparison of the Performance of MiSeq and HiSeq 2500 in a Microbiome Study

  • Na, Hee Sam;Yu, Yeuni;Kim, Si Yeong;Lee, Jae-Hyung;Chung, Jin
    • Microbiology and Biotechnology Letters
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    • v.48 no.4
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    • pp.574-581
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    • 2020
  • Next generation sequencing is commonly used to characterize the microbiome structure. MiSeq is commonly used to analyze the microbiome due to its relatively long read length. However, recently, Illumina introduced the 250x2 chip for HiSeq 2500. The purpose of this study was to compare the performance of MiSeq and HiSeq in the context of oral microbiome samples. The MiSeq Reagent Kit V3 and the HiSeq Rapid SBS Kit V2 were used for MiSeq and HiSeq 2500 analyses, respectively. Total read count, read quality score, relative bacterial abundance, community diversity, and relative abundance correlation were analyzed. HiSeq produced significantly more read sequences and assigned taxa compared to MiSeq. Conversely, community diversity was similar in the context of MiSeq and HiSeq. However, depending on the relative abundance, the correlation between the two platforms differed. The correlation between HiSeq and MiSeq sequencing data for highly abundant taxa (> 2%), low abundant taxa (2-0.2%), and rare taxa (0.2% >) was 0.994, 0.860, and 0.416, respectively. Therefore, HiSeq 2500 may also be compatible for microbiome studies. Importantly, the HiSeq platform may allow a high-resolution massive parallel sequencing for the detection of rare taxa.

Comparison of the MGISEQ-2000 and Illumina HiSeq 4000 sequencing platforms for RNA sequencing

  • Jeon, Sol A;Park, Jong Lyul;Kim, Jong-Hwan;Kim, Jeong Hwan;Kim, Yong Sung;Kim, Jin Cheon;Kim, Seon-Young
    • Genomics & Informatics
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    • v.17 no.3
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    • pp.32.1-32.6
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    • 2019
  • Currently, Illumina sequencers are the globally leading sequencing platform in the next-generation sequencing market. Recently, MGI Tech launched a series of new sequencers, including the MGISEQ-2000, which promise to deliver high-quality sequencing data faster and at lower prices than Illumina's sequencers. In this study, we compared the performance of two major sequencers (MGISEQ-2000 and HiSeq 4000) to test whether the MGISEQ-2000 sequencer delivers high-quality sequence data as suggested. We performed RNA sequencing of four human colon cancer samples with the two platforms, and compared the sequencing quality and expression values. The data produced from the MGISEQ-2000 and HiSeq 4000 showed high concordance, with Pearson correlation coefficients ranging from 0.98 to 0.99. Various quality control (QC) analyses showed that the MGISEQ-2000 data fulfilled the required QC measures. Our study suggests that the performance of the MGISEQ-2000 is comparable to that of the HiSeq 4000 and that the MGISEQ-2000 can be a useful platform for sequencing.

Draft genome sequence of Pelagicola sp. DSW4-44 isolated from seawater (해수에서 분리된 Pelagicola sp. DSW4-44의 초안 유전체 서열분석)

  • Oh, Ji-Sung;Roh, Dong-Hyun
    • Korean Journal of Microbiology
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    • v.55 no.3
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    • pp.283-285
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    • 2019
  • The draft genome sequencing for Pelagicola sp. DSW4-44 (= KCTC 62762 = KCCM 43261), isolated from deep seawater of East Sea in Korea, was performed using Illumina HiSeq platform. As a result, the draft genome was comprised of a total length of approximately 4.85 Mbp with G + C content of 54.3%, and included a total of 4,566 protein-coding genes, 3 rRNA genes, 48 tRNA genes, 3 non-coding RNA genes, and 67 pseudo genes. In the draft genome, the strain DSW4-44 contained genes involved in the nitrogen metabolism of dissimilatory nitrate reduction to ammonium (DNRA) and denitrification, which were not found other strains in the genus Pelagicola.

Draft genome sequence of Zhongshania marina DSW25-10T isolated from seawater (해수에서 분리된 Zhongshania marina DSW25-10T 의 유전체 서열분석)

  • Oh, Ji-Sung;Roh, Dong-Hyun
    • Korean Journal of Microbiology
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    • v.54 no.4
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    • pp.480-482
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    • 2018
  • The draft genome sequencing for Zhongshania marina $DSW25-10^T$, isolated from deep seawater of East Sea in Korea, was performed using Illumina HiSeq platform. As a result, the draft genome was comprised of a total length of approximately 4.08 Mbp with G + C content of 49.0%, and included a total of 3,702 protein-coding genes, 3 rRNA genes, 39 tRNA genes, 4 non-coding RNA genes, and 36 pseudogenes. In addition, the metabolic pathways of aliphatic and aromatic compounds were identified. In light of these metabolic pathways, Zhongshania marina $DSW25-10^T$ is expected to be a useful bioremediation resource.

Genome Survey and Microsatellite Marker Selection of Tegillarca granosa (꼬막(Tegillarca granosa)의 유전적 다양성 분석을 위한 드래프트 게놈분석과 마이크로새틀라이트 마커 발굴)

  • Kim, Jinmu;Lee, Seung Jae;Jo, Euna;Choi, Eunkyung;Kim, Hyeon Jin;Lee, Jung Sick;Park, Hyun
    • Journal of Marine Life Science
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    • v.6 no.1
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    • pp.38-46
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    • 2021
  • The blood clam, Tegillarca granosa, is economically important in marine bivalve and is used in fisheries industry among western Pacific Ocean Coasts especially in Korea, China, and Japan. The number of chromosomes in the blood clam is known as 2n=38, but the genome size and genetic information of the genome are not still clear. In order to predict the genomic size of the T. granosa, the in-silico analysis analysed the genomic size using short DNA sequence information obtained using the NGS Illumina HiSeq platform. As a result, the genomic size of T. granosa was estimated to be 770.61 Mb. Subsequently, a draft genome assembly was performed through the MaSuRCA assembler, and a simple sequence repeat (SSR) analysis was done by using the QDD pipeline. 43,944 SSRs were detected from the genome of T. granosa and 69.51% di-nucleotide, 16.68% trinucleotide, 12.96% tetra-nucleotide, 0.82% penta-nucleotide, and 0.03% hexa-nucleotide were consisted. 100 primer sets that could be used for genetic diversity studies were selected. In the future, this study will help identify the genetic diversity of T. granosa and population genetic studies, and further identify the classification of origin between homogenous groups.

Complete genome sequence of Comamonas sp. NLF-7-7 isolated from biofilter of wastewater treatment plant (폐수처리장의 바이오 필터로부터 분리된 Comamonas sp. NLF-7-7 균주의 유전체 염기서열 해독)

  • Kim, Dong-Hyun;Han, Kook-Il;Kwon, Hae Jun;Kim, Mi Gyeong;Kim, Young Guk;Choi, Doo Ho;Lee, Keun Chul;Suh, Min Kuk;Kim, Han Sol;Lee, Jung-Sook;Kim, Jong-Guk
    • Korean Journal of Microbiology
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    • v.55 no.3
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    • pp.309-312
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    • 2019
  • Comamonas sp. NLF-7-7 was isolated from biofilter of wastewater treatment plant. The whole-genome sequence of Comamonas sp. NLF-7-7 was analyzed using the PacBio RS II and Illumina HiSeqXten platform. The genome comprises a 3,333,437 bp chromosome with a G + C content of 68.04%, 3,197 total genes, 9 rRNA genes, and 49 tRNA genes. This genome contained pollutants degradation and floc forming genes such as sulfur oxidization pathway (SoxY, SoxZ, SoxA, and SoxB) and floc forming pathway (EpsG, EpsE, EpsF, EpsG, EpsL, and glycosyltransferase), respectively. The Comamonas sp. NLF-7-7 can be used to the purification of wastewater.

Toward Complete Bacterial Genome Sequencing Through the Combined Use of Multiple Next-Generation Sequencing Platforms

  • Jeong, Haeyoung;Lee, Dae-Hee;Ryu, Choong-Min;Park, Seung-Hwan
    • Journal of Microbiology and Biotechnology
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    • v.26 no.1
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    • pp.207-212
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    • 2016
  • PacBio's long-read sequencing technologies can be successfully used for a complete bacterial genome assembly using recently developed non-hybrid assemblers in the absence of second-generation, high-quality short reads. However, standardized procedures that take into account multiple pre-existing second-generation sequencing platforms are scarce. In addition to Illumina HiSeq and Ion Torrent PGM-based genome sequencing results derived from previous studies, we generated further sequencing data, including from the PacBio RS II platform, and applied various bioinformatics tools to obtain complete genome assemblies for five bacterial strains. Our approach revealed that the hierarchical genome assembly process (HGAP) non-hybrid assembler resulted in nearly complete assemblies at a moderate coverage of ~75x, but that different versions produced non-compatible results requiring post processing. The other two platforms further improved the PacBio assembly through scaffolding and a final error correction.

Metagenomic and Proteomic Analyses of a Mangrove Microbial Community Following Green Macroalgae Enteromorpha prolifera Degradation

  • Wu, Yijing;Zhao, Chao;Xiao, Zheng;Lin, Hetong;Ruan, Lingwei;Liu, Bin
    • Journal of Microbiology and Biotechnology
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    • v.26 no.12
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    • pp.2127-2137
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    • 2016
  • A mangrove microbial community was analyzed at the gene and protein levels using metagenomic and proteomic methods with the green macroalgae Enteromorpha prolifera as the substrate. Total DNA was sequenced on the Illumina HiSeq 2000 PE-100 platform. Two-dimensional gel electrophoresis in combination with liquid chromatography tandem mass spectrometry was used for proteomic analysis. The metagenomic data revealed that the orders Pseudomonadales, Rhizobiales, and Sphingomonadales were the most prevalent in the mangrove microbial community. By monitoring changes at the functional level, proteomic analyses detected ATP synthase and transporter proteins, which were expressed mainly by members of the phyla Proteobacteria and Bacteroidetes. Members of the phylum Proteobacteria expressed a high number of sugar transporters and demonstrated specialized and efficient digestion of various glycans. A few glycoside hydrolases were detected in members of the phylum Firmicutes, which appeared to be the main cellulose-degrading bacteria. This is the first report of multiple "omics" analysis of E. prolifera degradation. These results support the fact that key enzymes of glycoside hydrolase family were expressed in large quantities, indicating the high metabolic activity of the community.

The Complete Mitochondrial Genome of the Fourhorn Sculpin Triglopsis quadricornis (Perciformes, Cottidae) from Sirius Passet, North Greenland

  • Kim, Bo-Mi;Kihm, Ji-Hoon;Park, Tae-Yoon S.
    • Ocean and Polar Research
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    • v.43 no.4
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    • pp.371-374
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    • 2021
  • Triglopsis quadricornis Linnaeus, 1758 (Cottidae) is distributed in the Atlantic and Arctic and has four unique bony protuberances on its head. Here, we report the complete, circular, and annotated mitochondrial genome of T. quadricornis. The complete T. quadricornis mitochondrion was sequenced by high-throughput Illumina HiSeq platform. The sequences are 16,736 bp in size and contains 13 protein-coding genes (PCGs), 22 transfer RNA (tRNA) genes, a control region, and large and small ribosomal subunits. The overall genomic structure of T. quadricornis mitochondrion was conserved with the gene arrangement of Megalocottus and Myoxocephalus species, and phylogenetic analysis supports their sister relationships. Most PCGs consist of TAA or TAG as a termination codon, whereas COII, ND4, and CYTB have T-- as a stop codon. This complete mitochondrial DNA information of T. quadricornis will provide an essential genomic resource to elucidate the phylogenetic relationship and evolutionary history of the family Cottidae.

Effects of Heat-stress on Rumen Bacterial Diversity and Composition of Holstein Cows (고온 스트레스 영향에 따른 홀스타인종 젖소의 반추위내 미생물 균총 변화)

  • Kim, Dong Hyeon;Kim, Myung Hoo;Kim, Sang Bum;Ha, Seung Min;Son, Jun Kyu;Lee, Ji Hwan;Hur, Tai Young;Lee, Jae Yeong;Park, Ji Hoo;Choi, Hee Chul;Lee, Hyun Jeong;Park, Beom Young;Ki, Kwang Seok;Kim, Eun Tae
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.39 no.4
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    • pp.227-234
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    • 2019
  • This study was performed to investigate the effect of heat-stressed environment on rumen microbial diversity in Holstein cows. Rectal temperature and respiration rate were measured and rumen fluid was collected under normal environment (NE; Temperature humidity index (THI)=64.6) and heat-stressed environment (HE; THI=87.2) from 10 Holstein cows (60±17.7 months, 717±64.4 kg) fed on the basis of dairy feeding management in National Institute of Animal Science. The rumen bacteria diversity was analyzed by using the Illumina HiSeqTM 4000 platform. The rectal temperature and respiratory rate were increased by 1.5℃ and 53 breaths/min in HE compared to that in NE, respectively. In this study, HE exposure induced significant changes of ruminal microbe. At phylum level, Fibrobacteres were increased in HE. At genus level, Ruminococcaceae bacterium P7 and YAD3003, Butyrivibrio sp. AE2032, Erysipelotrichaceae bacterium NK3D112, Bifidobacterium pseudolongum, Lachnospiraceae bacterium FE2018, XBB2008, and AC2029, Eubacterium celulosolvens, Clostridium hathewayi, and Butyrivibrio hungatei were decreased in HE, while Choristoneura murinana nucleopolyhedrovirus, Calothrix parasitica, Nostoc sp. KVJ20, Anabaena sp. ATCC 33047, Fibrobacter sp. UWB13 and sp. UWB5, Lachnospiraceae bacterium G41, and Xanthomonas arboricola were increased in HE. In conclusion, HE might have an effect to change the rumen microbial community in Holstein cows.