• Title/Summary/Keyword: Sequencing depth

Search Result 50, Processing Time 0.026 seconds

Survey of the Applications of NGS to Whole-Genome Sequencing and Expression Profiling

  • Lim, Jong-Sung;Choi, Beom-Soon;Lee, Jeong-Soo;Shin, Chan-Seok;Yang, Tae-Jin;Rhee, Jae-Sung;Lee, Jae-Seong;Choi, Ik-Young
    • Genomics & Informatics
    • /
    • v.10 no.1
    • /
    • pp.1-8
    • /
    • 2012
  • Recently, the technologies of DNA sequence variation and gene expression profiling have been used widely as approaches in the expertise of genome biology and genetics. The application to genome study has been particularly developed with the introduction of the nextgeneration DNA sequencer (NGS) Roche/454 and Illumina/ Solexa systems, along with bioinformation analysis technologies of whole-genome $de$ $novo$ assembly, expression profiling, DNA variation discovery, and genotyping. Both massive whole-genome shotgun paired-end sequencing and mate paired-end sequencing data are important steps for constructing $de$ $novo$ assembly of novel genome sequencing data. It is necessary to have DNA sequence information from a multiplatform NGS with at least $2{\times}$ and $30{\times}$ depth sequence of genome coverage using Roche/454 and Illumina/Solexa, respectively, for effective an way of de novo assembly. Massive shortlength reading data from the Illumina/Solexa system is enough to discover DNA variation, resulting in reducing the cost of DNA sequencing. Whole-genome expression profile data are useful to approach genome system biology with quantification of expressed RNAs from a wholegenome transcriptome, depending on the tissue samples. The hybrid mRNA sequences from Rohce/454 and Illumina/Solexa are more powerful to find novel genes through $de$ $novo$ assembly in any whole-genome sequenced species. The $20{\times}$ and $50{\times}$ coverage of the estimated transcriptome sequences using Roche/454 and Illumina/Solexa, respectively, is effective to create novel expressed reference sequences. However, only an average $30{\times}$ coverage of a transcriptome with short read sequences of Illumina/Solexa is enough to check expression quantification, compared to the reference expressed sequence tag sequence.

Development of an SNP set for marker-assisted breeding based on the genotyping-by-sequencing of elite inbred lines in watermelon (수박 엘리트 계통의 GBS를 통한 마커이용 육종용 SNP 마커 개발)

  • Lee, Junewoo;Son, Beunggu;Choi, Youngwhan;Kang, Jumsoon;Lee, Youngjae;Je, Byoung Il;Park, Younghoon
    • Journal of Plant Biotechnology
    • /
    • v.45 no.3
    • /
    • pp.242-249
    • /
    • 2018
  • This study was conducted to develop an SNP set that can be useful for marker-assisted breeding (MAB) in watermelon (Citrullus. lanatus L) using Genotyping-by-sequencing (GBS) analysis of 20 commercial elite watermelon inbreds. The result of GBS showed that 77% of approximately 1.1 billion raw reads were mapped on the watermelon genome with an average mapping region of about 4,000 Kb, which indicated genome coverage of 2.3%. After the filtering process, a total of 2,670 SNPs with an average depth of 31.57 and the PIC (Polymorphic Information Content) value of 0.1~0.38 for 20 elite inbreds were obtained. Among those SNPs, 55 SNPs (5 SNPs per chromosome that are equally distributed on each chromosome) were selected. For the understanding genetic relationship of 20 elite inbreds, PCA (Principal Component Analysis) was carried out with 55 SNPs, which resulted in the classification of inbreds into 4 groups based on PC1 (52%) and PC2 (11%), thus causing differentiation between the inbreds. A similar classification pattern for PCA was observed from hierarchical clustering analysis. The SNP set developed in this study has the potential for application to cultivar identification, F1 seed purity test, and marker-assisted backcross (MABC) not only for 20 elite inbreds but also for diverse resources for watermelon breeding.

Phylogeny of Yeasts Isolated from the Flower of Aster spathulifolius Maxim. and Screening of Biosurfactant Producers (해국(Aster spathulifolius Maxim.)의 꽃에서 분리한 효모의 분자계통학적 분포해석 및 생물계면활성제 생산 균주의 스크리닝)

  • Kim, Jong-Shik;Kim, Dae-Shin
    • Korean Journal of Environmental Agriculture
    • /
    • v.37 no.4
    • /
    • pp.312-316
    • /
    • 2018
  • BACKGROUND: Yeast biotechnology finds applications in various industries. Hence, we sought to explore the yeasts associated with the flower of Aster spathulifolius Maxim. This study aimed to isolate yeasts from the flower of the plant and screen for biosurfactant-producing yeasts. METHODS AND RESULTS: We collected flowers of Aster spathulifolius Maxim. and performed pure isolation using four types of media. In total, 117 strains belonging to 4 genera, namely, Cryptococcus (75 strains), Aureobasidium pullulans (30 strains), Candida (11 strains), and Rhodotorula (1 strain), were isolated and identified by ITS sequencing. Upon in-depth analysis, Cryptococcus, the most dominant genus (75 strains) was categorized into the 'Unknown group'. Upon in-depth analysis of A. pullulans, we discovered the 'Unknown group I' (27 strains) and the 'Unknown group II' (2 strains), which have not been reported previously. Two A. pullulans isolates with potent surfactant activity were selected via the screening procedure. CONCLUSION: In this study, a total of 117 strains were isolated from the flower of Aster spathulifolius Maxim. In addition, two biosurfactant-producing yeasts were identified from among the isolated yeasts.

In Silico Metagenomes Mining to Discover Novel Esterases with Industrial Application by Sequential Search Strategies

  • Barriuso, Jorge;Jesus Martinez, Maria
    • Journal of Microbiology and Biotechnology
    • /
    • v.25 no.5
    • /
    • pp.732-737
    • /
    • 2015
  • We present here an in silico search of fungal sterol-esterase/lipase and bacterial depolymerase sequences from environmental metagenomes. Both enzyme types contain the α/β-hydrolase protein fold. Analysis of DNA conserved motifs, protein homology search, phylogenetic analysis, and protein 3D modeling have been used, and the efficiency of these screening strategies is discussed. The presence of bacterial genes in the metagenomes was higher than those from fungi, and the sequencing depth of the metagenomes seemed to be crucial to allow finding enough diversity of enzyme sequences. As a result, a novel putative PHA-depolymerase is described.

A Study on Cutting Tool Selection Techniques for Rough and Finish Turning Operations (선삭가공에서 황삭 및 정삭용 절삭공구선정방법에 관한 연구)

  • 김인호
    • Korean Journal of Computational Design and Engineering
    • /
    • v.3 no.4
    • /
    • pp.236-242
    • /
    • 1998
  • This paper presents a development of computer aided cutting tool selection techniques for rough and finish turning operations. The developed system,. which is one of important activities for computer aided operation planning, firstly implements operation sequencing. Then, from relations of the size of machined area, recommended finishing allowance and maximum depth of cut, a main machining method is selected, a number of cut is calculated, cutting tools including toolholders and inserts are selected, and values for cutting parameters are determined. A cutting tool selection procedure is proposed for toolholders and inserts of ISO code in rough cutting, and some important parameters such as holder style, tool approach angle, tool function and its direction are described in detail. In order to demonstrate the validity of the system a case study is performed.

  • PDF

Dietary turmeric (Curcuma longa L.) supplementation improves growth performance, short-chain fatty acid production, and modulates bacterial composition of weaned piglets

  • Recharla, Neeraja;Balasubramanian, Balamuralikrishnan;Song, Minho;Puligundla, Pradeep;Kim, Soo-ki;Jeong, Jin Young;Park, Sungkwon
    • Journal of Animal Science and Technology
    • /
    • v.63 no.3
    • /
    • pp.575-592
    • /
    • 2021
  • In livestock nutrition, natural feed additives are gaining increased attention as alternatives to antibiotic growth promoters to improve animal performance. This study investigated the effects of dietary turmeric supplementation on the growth performance and gut health of weaned piglets. A total of 48 weaned piglets (Duroc × [Landrace × Yorkshire]) were used in a 6-week feeding trial. All piglets were allotted to two dietary treatments: corn-soybean meal basal diet without turmeric (control) and with 1% weight per weight (w/w) turmeric powder (turmeric). The results showed that dietary inclusion of turmeric with the basal diet improved final body weight and total average daily gain (p < 0.05). The concentrations of short-chain fatty acids in the fecal samples, including acetic, butyric, and propionic acids, were higher in the turmeric group (p < 0.05). The villus height-to-crypt depth ratio was higher in the ileum of turmeric-fed piglets (p = 0.04). The 16S rRNA gene sequencing of fecal microbiota indicated that, at the phylum level, Firmicutes and Bacteroidetes were the most predominant taxa in all fecal samples. Bacteroidetes were significantly decreased in the turmeric group compared to the control group (p = 0.021). At the genus level, turmeric showed a decreased abundance of Prevotella (p = 0.021) and an increasing trend of Lactobacillus (p = 0.083). Among the total detected species, nine bacterial species showed significant differences between the two groups. The results of this study indicated that turmeric altered the gut microbiota and shortchain fatty acid production. This suggests that turmeric could be used as a potential alternative growth promoter for piglets.

Phylogeny of Marine Yeasts Isolated from Coastal Seawater in the East Sea of Korea (동해 연안해역에서 분리한 해양효모의 분자계통학적 해석)

  • Chin, Il-Seok;Kim, Yong-Hwan;Yun, Won-Kap;Park, Nyun-Ho;Kim, Jong-Shik
    • Korean Journal of Environmental Agriculture
    • /
    • v.36 no.2
    • /
    • pp.129-134
    • /
    • 2017
  • BACKGROUND: Yeasts are used in a variety of industries. However, most industries are biased toward Saccharomyces cerevisiae ; so we sought to explore non-conventional yeasts (NCY). This study aimed to isolate yeasts from seawater collected from the East Sea of Korea and to analyze the NCY. METHODS AND RESULTS: We first collected seawater and performed pure isolation using four kinds of medium (GPY, DOB + CSM, DG18, and SCG). In total, 314 strains and 17 genera were isolated by ITS sequencing, including Aureobasidum pullulans (236 strains), Cryptococcus (19 strains), Cystobasidium (18 strains), and Rhodotorula (9 strains). Upon in-depth analysis, A. pullulans, the most dominant genus (236 strains), was divided into Group II (147 strains), Unknown I (8 strains), and Unknown II (49 strains). CONCLUSION: In this study, a total of 314 strains were isolated from seawater; many of these yeasts have been found and reported in seawater previously. In-depth analysis of A. pullulans, showed the dominance of Group I (21 strains) and Group II (147 strains) We also discovered Unknown I (8 strains) and Unknown II (49 strains), which have not been reported previously.

Community Structure, Diversity, and Vertical Distribution of Archaea Revealed by 16S rRNA Gene Analysis in the Deep Sea Sediment of the Ulleung Basin, East Sea (16S rRNA 유전자 분석방법을 이용한 동해 울릉분지 심해 퇴적물 내 고세균 군집 구조 및 다양성의 수직분포 특성연구)

  • Kim, Bo-Bae;Cho, Hye-Youn;Hyun, Jung-Ho
    • Ocean and Polar Research
    • /
    • v.32 no.3
    • /
    • pp.309-319
    • /
    • 2010
  • To assess community structure and diversity of archaea, a clone sequencing analysis based on an archaeal 16S rRNA gene was conducted at three sediment depths of the continental slope and Ulleung Basin in the East Sea. A total of 311 and 342 clones were sequenced at the slope and basin sites, respectively. Marine Group I, which is known as the ammonia oxidizers, appeared to predominate in the surface sediment of both sites (97.3% at slope, 88.5% at basin). In the anoxic subsurface sediment of the slope and basin, the predominant archaeal group differed noticeably. Marine Benthic Group B dominated in the subsurface sediment of the slope. Marine Benthic Group D and Miscellaneous Crenarchaeotal Group were the second largest archaeal group at 8-9 cm and 18-19 cm depth, respectively. Marine Benthic Group C of Crenarchaeota occupied the highest proportion by accounting for more than 60% of total clones in the subsurface sediments of the basin site. While archaeal groups that use metal oxide as an electron acceptor were relatively more abundant at the basin sites with manganese (Mn) oxide-enriched surface sediment, archaeal groups related to the sulfur cycle were more abundant in the sulfidogenic sediments of the slope. Overall results indicate that archaeal communities in the Ulleung Basin show clear spatial variation with depth and sites according to geochemical properties the sediment. Archaeal communities also seem to play a significant role in the biogeochemical carbon (C), nitrogen (N), sulfur (S), and metal cycles at each site.

Long non-coding RNAs in Sus scrofa ileum under starvation stress

  • Wang, Shu;Ma, Yi Jia;Li, Yong Shi;Ge, Xu Sheng;Lu, Chang;Cai, Chun Bo;Yang, Yang;Zhao, Yan;Liang, Guo Ming;Guo, Xiao Hong;Cao, Guo Qing;Li, Bu Gao;Gao, Peng Fei
    • Animal Bioscience
    • /
    • v.35 no.7
    • /
    • pp.975-988
    • /
    • 2022
  • Objective: In this study, we aimed to identify long non-coding RNAs (lncRNAs) that play important roles in starvation stress, analyze their functions, and discover potential molecular targets to alleviate starvation stress to provide a theoretical reference for subsequent in-depth research. Methods: We generated a piglet starvation stress animal model. Nine Yorkshire weaned piglets were randomly divided into a long-term starvation stress group (starved for 72 h), short-term starvation stress group (starved for 48 h), and the control group. LncRNA libraries were constructed using high-throughput sequencing of piglet ileums. Results: We obtained 11,792 lncRNAs, among which, 2,500 lncRNAs were novel. In total, 509 differentially expressed (DE)lncRNAs were identified in this study. Target genes of DElncRNAs were predicted via cis and trans interactions, and functional and pathway analyses were performed. Gene ontology functions and Kyoto encyclopedia of genes and genomes analysis revealed that lncRNA-targeted genes mainly participated in metabolic pathways, cellular processes, immune system processes, digestive systems, and transport activities. To reveal the mechanism underlying starvation stress, the interaction network between lncRNAs and their targets was constructed based on 26 DElncRNAs and 72 DEmRNAs. We performed an interaction network analysis of 121 DElncRNA-DEmRNA pairs with a Pearson correlation coefficient greater than 0.99. Conclusion: We found that MSTRG.19894.13, MSTRG.16726.3, and MSTRG.12176.1 might play important roles in starvation stress. This study not only generated a library of enriched lncRNAs in piglets, but its outcomes also provide a strong foundation to screen key lncRNAs involved in starvation stress and a reference for subsequent in-depth research.

Bacterial Community Structure and Diversity Using 16S rDNA Analysis in the Intertidal Sediment of Ganghwa Island (16S rDNA 분석을 이용한 강화도 장화리 갯벌 퇴적물 내 미생물 군집구조 및 다양성)

  • Cho Hye Youn;Lee Jung-Hyun;Hyun Jung-Ho
    • Korean Journal of Microbiology
    • /
    • v.40 no.3
    • /
    • pp.189-198
    • /
    • 2004
  • T-RFLP analysis and clone sequencing analysis based on bacterial 16S rDNA were conducted to assess bacterial community structure and diversity in two layers (0-1cm, 6-7cm depth) of the sediment from Janghwari intertidal flat in Ganghwa Island. The results of T-RFLP (terminal-restriction fragment length polymorphism) analysis using restriction enzyme HhaI showed that the T-RFs of various size ($60{\pm}2$) bp-($667{\pm}2$) bp) appeared evenly at the surface sediments but two T-RFs with 60(${\pm}2$)bp and 93 (${\pm}2$)bp predominated at 6-7cm depth. Analysis of partial sequences for 172 clones revealed that 98% of the clones were not matched with the sequences of cultured bacteria strains in the GenBank (${\geq}similarity$ 98%), and approximately 86% of them were classified as different phylotypes. Most clones belonged to $\alpha$-, $\gamma$-, and $\delta$-Proteobacteria, Acidobacteria/Holophaga and green nonsulfur bacteria group. Proteobacteria group occupied the highest proportion in both layers (69% at 0-1cm depth and 46% at 6-7cm depth). $\gamma$-Proteobacteria and $\delta$-Proteobacteria that are associated with oxidation and reduction of sulfur compounds were appeared to be dominant, and comprised 21.5% and 15.7% of total clones, respectively. Overall results indicated that extremely diverse bacterial groups were inhabiting in the sediment of Ganghwa intertidal flat, and bacterial communities associated with the behaviour of sulfur seemed to playa significant role in the biogeochemical environment in this anoxic sediment.