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      Easymap: A User-Friendly Software Package for Rapid Mapping-by-Sequencing of Point Mutations and Large Insertions

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          Abstract

          Mapping-by-sequencing strategies combine next-generation sequencing (NGS) with classical linkage analysis, allowing rapid identification of the causal mutations of the phenotypes exhibited by mutants isolated in a genetic screen. Computer programs that analyze NGS data obtained from a mapping population of individuals derived from a mutant of interest to identify a causal mutation are available; however, the installation and usage of such programs requires bioinformatic skills, modifying or combining pieces of existing software, or purchasing licenses. To ease this process, we developed Easymap, an open-source program that simplifies the data analysis workflows from raw NGS reads to candidate mutations. Easymap can perform bulked segregant mapping of point mutations induced by ethyl methanesulfonate (EMS) with DNA-seq or RNA-seq datasets, as well as tagged-sequence mapping for large insertions, such as transposons or T-DNAs. The mapping analyses implemented in Easymap have been validated with experimental and simulated datasets from different plant and animal model species. Easymap was designed to be accessible to all users regardless of their bioinformatics skills by implementing a user-friendly graphical interface, a simple universal installation script, and detailed mapping reports, including informative images and complementary data for assessment of the mapping results. Easymap is available at http://genetics.edu.umh.es/resources/easymap; its Quickstart Installation Guide details the recommended procedure for installation.

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          Most cited references51

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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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            Fast gapped-read alignment with Bowtie 2.

            As the rate of sequencing increases, greater throughput is demanded from read aligners. The full-text minute index is often used to make alignment very fast and memory-efficient, but the approach is ill-suited to finding longer, gapped alignments. Bowtie 2 combines the strengths of the full-text minute index with the flexibility and speed of hardware-accelerated dynamic programming algorithms to achieve a combination of high speed, sensitivity and accuracy.
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              Graph-based genome alignment and genotyping with HISAT2 and HISAT-genotype

              Rapid advances in next-generation sequencing technologies have dramatically changed our ability to perform genome-scale analyses. The human reference genome used for most genomic analyses represents only a small number of individuals, limiting its usefulness for genotyping. We designed a novel method, HISAT2, for representing and searching an expanded model of the human reference genome, in which a large catalogue of known genomic variants and haplotypes is incorporated into the data structure used for searching and alignment. This strategy for representing a population of genomes, along with a fast and memory-efficient search algorithm, enables more detailed and accurate variant analyses than previous methods. We demonstrate two initial applications of HISAT2: HLA typing, a critical need in human organ transplantation, and DNA fingerprinting, widely used in forensics. These applications are part of HISAT-genotype, with performance not only surpassing earlier computational methods, but matching or exceeding the accuracy of laboratory-based assays.
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                Author and article information

                Contributors
                Journal
                Front Plant Sci
                Front Plant Sci
                Front. Plant Sci.
                Frontiers in Plant Science
                Frontiers Media S.A.
                1664-462X
                07 May 2021
                2021
                : 12
                : 655286
                Affiliations
                Instituto de Bioingeniería, Universidad Miguel Hernández de Elche , Elche, Spain
                Author notes

                Edited by: Marcos Egea-Cortines, Universidad Politécnica de Cartagena, Spain

                Reviewed by: Unai López De Heredia, Polytechnic University of Madrid, Spain; Hiroyuki Koga, The University of Tokyo, Japan

                *Correspondence: José Luis Micol, jlmicol@ 123456umh.es

                These authors have contributed equally to this work

                Present address: David Wilson-Sánchez, Department of Comparative Development and Genetics, Max Planck Institute for Plant Breeding Research, Cologne, Germany; Sergio Andreu-Sánchez, Department of Pediatrics and Genetics, University of Groningen, Groningen, Netherlands

                This article was submitted to Technical Advances in Plant Science, a section of the journal Frontiers in Plant Science

                Article
                10.3389/fpls.2021.655286
                8143052
                8e34b1f1-7214-40f0-80e4-bd96e987d083
                Copyright © 2021 Lup, Wilson-Sánchez, Andreu-Sánchez and Micol.

                This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

                History
                : 18 January 2021
                : 18 March 2021
                Page count
                Figures: 3, Tables: 4, Equations: 0, References: 51, Pages: 10, Words: 0
                Funding
                Funded by: Ministerio de Ciencia e Innovación 10.13039/501100004837
                Funded by: Generalitat Valenciana 10.13039/501100003359
                Categories
                Plant Science
                Methods

                Plant science & Botany
                bioinformatics,ngs,mapping-by-sequencing,candidate mutations,forward genetics,linkage analysis mapping,bulked segregant analysis

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