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      Genomic and transcriptomic variation defines the chromosome-scale assembly of Haemonchus contortus, a model gastrointestinal worm

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          Abstract

          Haemonchus contortus is a globally distributed and economically important gastrointestinal pathogen of small ruminants and has become a key nematode model for studying anthelmintic resistance and other parasite-specific traits among a wider group of parasites including major human pathogens. Here, we report using PacBio long-read and OpGen and 10X Genomics long-molecule methods to generate a highly contiguous 283.4 Mbp chromosome-scale genome assembly including a resolved sex chromosome for the MHco3(ISE).N1 isolate. We show a remarkable pattern of conservation of chromosome content with Caenorhabditis elegans, but almost no conservation of gene order. Short and long-read transcriptome sequencing allowed us to define coordinated transcriptional regulation throughout the parasite’s life cycle and refine our understanding of cis- and trans-splicing. Finally, we provide a comprehensive picture of chromosome-wide genetic diversity both within a single isolate and globally. These data provide a high-quality comparison for understanding the evolution and genomics of Caenorhabditis and other nematodes and extend the experimental tractability of this model parasitic nematode in understanding helminth biology, drug discovery and vaccine development, as well as important adaptive traits such as drug resistance.

          Abstract

          Stephen Doyle et al. report the chromosome-scale genome assembly and transcriptome sequence data of Haemonchus contortus, a key parasitic nematode model. These data show nearly complete conservation of chromosome content with C. elegans and brings insight into transcriptional regulation and chromosome-wide genetic diversity in this important pathogen.

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          The Sequence Alignment/Map format and SAMtools

          Summary: The Sequence Alignment/Map (SAM) format is a generic alignment format for storing read alignments against reference sequences, supporting short and long reads (up to 128 Mbp) produced by different sequencing platforms. It is flexible in style, compact in size, efficient in random access and is the format in which alignments from the 1000 Genomes Project are released. SAMtools implements various utilities for post-processing alignments in the SAM format, such as indexing, variant caller and alignment viewer, and thus provides universal tools for processing read alignments. Availability: http://samtools.sourceforge.net Contact: rd@sanger.ac.uk
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            STAR: ultrafast universal RNA-seq aligner.

            Accurate alignment of high-throughput RNA-seq data is a challenging and yet unsolved problem because of the non-contiguous transcript structure, relatively short read lengths and constantly increasing throughput of the sequencing technologies. Currently available RNA-seq aligners suffer from high mapping error rates, low mapping speed, read length limitation and mapping biases. To align our large (>80 billon reads) ENCODE Transcriptome RNA-seq dataset, we developed the Spliced Transcripts Alignment to a Reference (STAR) software based on a previously undescribed RNA-seq alignment algorithm that uses sequential maximum mappable seed search in uncompressed suffix arrays followed by seed clustering and stitching procedure. STAR outperforms other aligners by a factor of >50 in mapping speed, aligning to the human genome 550 million 2 × 76 bp paired-end reads per hour on a modest 12-core server, while at the same time improving alignment sensitivity and precision. In addition to unbiased de novo detection of canonical junctions, STAR can discover non-canonical splices and chimeric (fusion) transcripts, and is also capable of mapping full-length RNA sequences. Using Roche 454 sequencing of reverse transcription polymerase chain reaction amplicons, we experimentally validated 1960 novel intergenic splice junctions with an 80-90% success rate, corroborating the high precision of the STAR mapping strategy. STAR is implemented as a standalone C++ code. STAR is free open source software distributed under GPLv3 license and can be downloaded from http://code.google.com/p/rna-star/.
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              Cutadapt removes adapter sequences from high-throughput sequencing reads

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                Author and article information

                Contributors
                stephen.doyle@sanger.ac.uk
                jc17@sanger.ac.uk
                Journal
                Commun Biol
                Commun Biol
                Communications Biology
                Nature Publishing Group UK (London )
                2399-3642
                9 November 2020
                9 November 2020
                2020
                : 3
                : 656
                Affiliations
                [1 ]GRID grid.10306.34, ISNI 0000 0004 0606 5382, Wellcome Sanger Institute, ; Hinxton, Cambridgeshire CB10 1SA UK
                [2 ]GRID grid.8756.c, ISNI 0000 0001 2193 314X, Institute of Biodiversity Animal Health and Comparative Medicine, College of Medical, Veterinary and Life Sciences, University of Glasgow, ; Garscube Campus Glasgow, G61 1QH UK
                [3 ]GRID grid.420013.4, ISNI 0000 0001 2186 0964, Moredun Research Institute, Pentlands Science Park, ; Bush Loan Penicuik, EH26 0PZ UK
                [4 ]GRID grid.14709.3b, ISNI 0000 0004 1936 8649, Institute of Parasitology, McGill University, ; 21111 Lakeshore Road, Sainte Anne-de-Bellevue, QC H9X3V9 Canada
                [5 ]GRID grid.4305.2, ISNI 0000 0004 1936 7988, Royal (Dick) School of Veterinary Studies, University of Edinburgh, ; Edinburgh, EH25 9RG UK
                [6 ]GRID grid.225360.0, ISNI 0000 0000 9709 7726, European Molecular Biology Laboratory, European Bioinformatics Institute, ; Hinxton, Cambridgeshire CB10 1SA UK
                [7 ]GRID grid.22072.35, ISNI 0000 0004 1936 7697, Department of Comparative Biology and Experimental Medicine, Faculty of Veterinary Medicine, University of Calgary, ; Calgary, AB Canada
                [8 ]INRAE - U. Tours, UMR 1282 ISP Infectiologie et Santé Publique, Centre de recherche Val de Loire, Nouzilly, France
                [9 ]GRID grid.412967.f, University of Veterinary and Animal Sciences, ; Lahore, 54600 Pakistan
                Author information
                http://orcid.org/0000-0001-9167-7532
                http://orcid.org/0000-0002-4805-9058
                http://orcid.org/0000-0002-6191-0759
                http://orcid.org/0000-0001-6677-2319
                http://orcid.org/0000-0003-2391-9573
                http://orcid.org/0000-0002-3116-744X
                http://orcid.org/0000-0002-1935-834X
                http://orcid.org/0000-0002-9581-0377
                http://orcid.org/0000-0002-6787-4699
                Article
                1377
                10.1038/s42003-020-01377-3
                7652881
                33168940
                9cbf14b4-3207-4378-9c25-4a40ed21722b
                © The Author(s) 2020

                Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

                History
                : 5 March 2020
                : 14 October 2020
                Funding
                Funded by: FundRef https://doi.org/10.13039/100004440, Wellcome Trust (Wellcome);
                Award ID: WT206194
                Award ID: 067811
                Award Recipient :
                Funded by: FundRef https://doi.org/10.13039/501100000268, RCUK | Biotechnology and Biological Sciences Research Council (BBSRC);
                Award ID: BB/M003949/1
                Award ID: BB/P024610/1
                Award ID: BB/K020048/1
                Award Recipient :
                Categories
                Article
                Custom metadata
                © The Author(s) 2020

                non-model organisms,genome informatics,parasitic infection,comparative genomics,population genetics

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