• 제목/요약/키워드: sequencing analysis

검색결과 1,937건 처리시간 0.026초

ChIP-seq Analysis of Histone H3K27ac and H3K27me3 Showing Different Distribution Patterns in Chromatin

  • Kang, Jin;Kim, AeRi
    • 대한의생명과학회지
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    • 제28권2호
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    • pp.109-119
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    • 2022
  • Histone proteins can be modified by the addition of acetyl group or methyl group to specific amino acids. The modifications have different distribution patterns in chromatin. Recently, histone modifications are studied based on ChIP-seq data, which requires reasonable analysis of sequencing data depending on their distribution patterns. Here we have analyzed histone H3K27ac and H3K27me3 ChIP-seq data and it showed that the H3K27ac is enriched at narrow regions while H3K27me3 distributes broadly. To properly analyze the ChIP-seq data, we called peaks for H3K27ac and H3K27me3 using MACS2 (narrow option and broad option) and SICER methods, and compared propriety of the peaks using signal-to-background ratio. As results, H3K27ac-enriched regions were well identified by both methods while H3K27me3 peaks were properly identified by SICER, which indicates that peak calling method is more critical for histone modifications distributed broadly. When ChIP-seq data were compared in different sequencing depth (15, 30, 60, 120 M), high sequencing depth caused high false-positive rate in H3K27ac peak calling, but it reflected more properly the broad distribution pattern of H3K27me3. These results suggest that sequencing depth affects peak calling from ChIP-seq data and high sequencing depth is required for H3K27me3. Taken together, peak calling tool and sequencing depth should be chosen depending on the distribution pattern of histone modification in ChIP-seq analysis.

합류하는 두 항공기간 도착순서 결정에 대한 로지스틱회귀 예측 모형 (Prediction Model with a Logistic Regression of Sequencing Two Arrival Flows)

  • 정소연;이금진
    • 한국항공운항학회지
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    • 제23권4호
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    • pp.42-48
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    • 2015
  • This paper has its purpose on constructing a prediction model of the arrival sequencing strategy which reflects the actual sequencing patterns of air traffic controllers. As the first step, we analyzed a pair-wise sequencing of two aircraft entering TMA from different entering points. Based on the historical trajectory data, several traffic factors such as time, speed and traffic density were examined for the model. With statistically significant factors, we constructed a prediction model of arrival sequencing through a binary logistic regression analysis. With the estimated coefficients, the performance of the model was conducted through a cross validation.

Development of an efficient genotyping-by-sequencing (GBS) library construction method for genomic analysis of grapevine

  • Jang, Hyun A;Oh, Sang-Keun
    • 농업과학연구
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    • 제44권4호
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    • pp.495-503
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    • 2017
  • Genotyping-by-sequencing (GBS) is an outstanding technology for genotyping and single nucleotide polymorphism (SNP) discovery compared to next generation sequencing (NGS) because it can save time when analyzing large-scale samples and carries a low cost per sample. Recently, studies using GBS have been conducted on major crops and, to a greater extent, on fruit crops. However, many researchers have some problems due to low GBS efficiency resulting from low quality GBS libraries. To overcome this limitation, we developed an efficient GBS library construction method that regulates important conditions such as restriction enzymes (RE) digestion and a PCR procedure for grapevine. For RE digestion, DNA samples are digested with ApeKI (3.6U) at $75^{\circ}C$ for 5 hours and adapters are ligated to the ends of gDNA products. To produce suitable PCR fragments for sequencing, we modified the PCR amplification conditions; temperature cycling consisted of $72^{\circ}C$ (5 min), $98^{\circ}C$ (30 s), followed by 16 cycles of $98^{\circ}C$ (30 s), $65^{\circ}C$ (30 s), $72^{\circ}C$ (20 s) with a final extension step. As a result, we had obtained optimal library construct sizes (200 to 400 bp) for GBS analysis. Furthermore, it not only increased the mapping efficiency by approximately 10.17% compared to the previous method, but also produced mapped reads which were distributed equally on the19 chromosomes in the grape genome. Therefore, we suggest that this system can be used for various fruit crops and is expected to increase the efficiency of various genomic analysis performed.

Trimming conditions for DADA2 analysis in QIIME2 platform

  • Lee, Seo-Young;Yu, Yeuni;Chung, Jin;Na, Hee Sam
    • International Journal of Oral Biology
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    • 제46권3호
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    • pp.146-153
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    • 2021
  • Accurate identification of microbes facilitates the prediction, prevention, and treatment of human diseases. To increase the accuracy of microbiome data analysis, a long region of the 16S rRNA is commonly sequenced via paired-end sequencing. In paired-end sequencing, a sufficient length of overlapping region is required for effective joining of the reads, and high-quality sequencing reads are needed at the overlapping region. Trimming sequences at the reads distal to a point where sequencing quality drops below a specific threshold enhance the joining process. In this study, we examined the effect of trimming conditions on the number of reads that remained after quality control and chimera removal in the Illumina paired-end reads of the V3-V4 hypervariable region. We also examined the alpha diversity and taxa assigned by each trimming condition. Optimum quality trimming increased the number of good reads and assigned more number of operational taxonomy units. The pre-analysis trimming step has a great influence on further microbiome analysis, and optimized trimming conditions should be applied for Divisive Amplicon Denoising Algorithm 2 analysis in QIIME2 platform.

Annotation of Genes Having Candidate Somatic Mutations in Acute Myeloid Leukemia with Whole-Exome Sequencing Using Concept Lattice Analysis

  • Lee, Kye Hwa;Lim, Jae Hyeun;Kim, Ju Han
    • Genomics & Informatics
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    • 제11권1호
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    • pp.38-45
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    • 2013
  • In cancer genome studies, the annotation of newly detected oncogene/tumor suppressor gene candidates is a challenging process. We propose using concept lattice analysis for the annotation and interpretation of genes having candidate somatic mutations in whole-exome sequencing in acute myeloid leukemia (AML). We selected 45 highly mutated genes with whole-exome sequencing in 10 normal matched samples of the AML-M2 subtype. To evaluate these genes, we performed concept lattice analysis and annotated these genes with existing knowledge databases.

ChIP-seq 라이브러리 제작 및 Galaxy 플랫폼을 이용한 NGS 데이터 분석 (ChIP-seq Library Preparation and NGS Data Analysis Using the Galaxy Platform)

  • 강유진;강진;김예운;김애리
    • 생명과학회지
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    • 제31권4호
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    • pp.410-417
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    • 2021
  • NGS (Next-generation sequencing), 즉 차세대염기서열분석은 유전체 수준의 방대한 DNA를 작은 절편으로 만들어서 그 절편들의 염기서열들을 동시에 읽어내는 기법이다. 현재 다양한 생명체의 유전체 염기서열 분석부터 cDNA (complementary DNA)나 ChIPed DNA (chromatin immunoprecipitated DNA)를 분석하는데 이 NGS 기법을 사용하고 있으며, 이 때 얻어진 데이터를 적절히 처리하고 분석하는 일은 생물학적으로 유의미한 결과를 얻기 위하여 중요하다. 하지만 대용량 데이터의 저장 및 활용, 그리고 컴퓨터 프로그래밍 바탕의 데이터 분석은 실험을 수행하는 일반 생물학자들에게 어려운 일이다. Galaxy 플랫폼은 다양한 NGS 데이터 분석 tool을 무료로 제공하는 웹 서비스이며, 생물정보학이나 프로그래밍에 대한 전문지식이 없는 연구자들에게 웹 브라우저만을 이용하여 데이터를 분석할 수 있는 환경을 제공한다. 본 논문에서는 ChIP-seq (chromatin immunoprecipitation-sequencing) 수행을 위한 라이브러리 제작 과정 및 Galaxy 플랫폼을 이용한 ChIP-seq 데이터 분석 과정을 설명하고, K562 세포주에서 수행한 히스톤 H3K4me1 ChIP-seq 결과가 public 데이터와 일치함을 보여준다. 따라서 Galaxy 플랫폼을 활용한 NGS 데이터 분석은 생물정보학에 대한 손쉬운 접근 방법을 제공할 것으로 기대된다.

COEX-Seq: Convert a Variety of Measurements of Gene Expression in RNA-Seq

  • Kim, Sang Cheol;Yu, Donghyeon;Cho, Seong Beom
    • Genomics & Informatics
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    • 제16권4호
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    • pp.36.1-36.3
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    • 2018
  • Next generation sequencing (NGS), a high-throughput DNA sequencing technology, is widely used for molecular biological studies. In NGS, RNA-sequencing (RNA-Seq), which is a short-read massively parallel sequencing, is a major quantitative transcriptome tool for different transcriptome studies. To utilize the RNA-Seq data, various quantification and analysis methods have been developed to solve specific research goals, including identification of differentially expressed genes and detection of novel transcripts. Because of the accumulation of RNA-Seq data in the public databases, there is a demand for integrative analysis. However, the available RNA-Seq data are stored in different formats such as read count, transcripts per million, and fragments per kilobase million. This hinders the integrative analysis of the RNA-Seq data. To solve this problem, we have developed a web-based application using Shiny, COEX-seq (Convert a Variety of Measurements of Gene Expression in RNA-Seq) that easily converts data in a variety of measurement formats of gene expression used in most bioinformatic tools for RNA-Seq. It provides a workflow that includes loading data set, selecting measurement formats of gene expression, and identifying gene names. COEX-seq is freely available for academic purposes and can be run on Windows, Mac OS, and Linux operating systems. Source code, sample data sets, and supplementary documentation are available as well.

고체상 나노구멍을 이용한 DNA 염기서열 분석기술 (DNA Sequencing Analysis Technique by Using Solid-State Nanopore)

  • 김태헌;박정호
    • 센서학회지
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    • 제21권5호
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    • pp.359-366
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    • 2012
  • Nanopore DNA sequencing is an emerging and promising technique that can potentially realize the goal of a low-cost and high-throughput method for analyzing human genome. Especially, solid-state nanopores have relatively high mechanical stability, simple surface modification, and facile fabrication process without the need for labeling or amplification of PCR (polymerized chain reaction) in DNA sequencing. For these advantages of solid-sate nanopores, the use of solid-state nanopores has been extensively considered for developing a next generation DNA sequencing technology. Solid-state nanopore sequencing technique can determine and count charged molecules such as single-stranded DNA, double-stranded DNA, or RNA when they are driven to pass through a membrane nanopore between two electrolytes of cis-trans chambers with applied bias voltage by measuring the ionic current which varies due to the existence of the charged particles in the nanopore. Recently, many researchers have suggested that nanopore-based sensors can be competitive with other third-generation DNA sequencing technologies, and may be able to rapidly and reliably sequence the human genome for under $1,000.

Cervus elaphus 종의 sequencing과 BLAST search에 의한 감별 (Identification of Cervus elaphus Species by Sequencing Analysis and BLAST Search)

  • 서정철;김민정;이찬;임강현
    • 대한본초학회지
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    • 제21권2호
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    • pp.129-133
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    • 2006
  • Objectives : Cervus elaphus species are some of the most medicinally important genera in the Oriental medicine. This study was performed to determine if Cenvus elaphus species could be identified by sequencing analysis and to verify Basic Local Alignment Search Tool (BLAST) search, which was used to assess genetic identification. Methods : The DNAs of Cervus elaphus species were extracted, amplified by PCR, and sequenced. The DNAs of Cervus species were identified by BLAST search in website. Results : By BLAST search one of Cervus elaphus species was identified as Cervus elaphussibericus but the other was identified as Cervus elaphus nelsoni. This work showed that identification can efficiently be performed by BLAST search. Conclusion : These results suggest that sequencing following BLAST search might be able to provide the identification of Cervus elaphus species.

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Application of genotyping-by-sequencing (GBS) in plant genome using bioinformatics pipeline

  • Lee, Yun Gyeong;Kang, Chon-Sik;Kim, Changsoo
    • 한국작물학회:학술대회논문집
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    • 한국작물학회 2017년도 9th Asian Crop Science Association conference
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    • pp.58-58
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    • 2017
  • The advent of next generation sequencing technology has elicited plenty of sequencing data available in agriculturally relevant plant species. For most crop species, it is too expensive to obtain the whole genome sequence data with sufficient coverage. Thus, many approaches have been developed to bring down the cost of NGS. Genotyping-by-sequencing (GBS) is a cost-effective genotyping method for complex genetic populations. GBS can be used for the analysis of genomic selection (GS), genome-wide association study (GWAS) and constructing haplotype and genetic linkage maps in a variety of plant species. For efficiently dealing with plant GBS data, the TASSEL-GBS pipeline is one of the most popular choices for many researchers. TASSEL-GBS is JAVA based a software package to obtain genotyping data from raw GBS sequences. Here, we describe application of GBS and bioinformatics pipeline of TASSEL-GBS for analyzing plant genetics data.

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