Abstract

When sequencing eukaryotic genomes, linkage maps are indispensable for building scaffolds to assemble and/or to validate chromosomes. However, current approaches to constructing linkage maps are limited by marker density and cost-effectiveness, especially for wild organisms. We have now devised a new strategy based on artificially generated hybrid organisms to acquire ultrahigh-density genomic markers at reduced cost and build highly accurate linkage maps. We have also developed the novel analysis pipeline Scaffold Extender with Low Depth Linkage Analysis (SELDLA) for data processing to generate linkage maps and draft genomes. Using SELDLA, linkage maps and improved genomes for two species of pufferfish, Takifugu rubripes and Takifugu stictonotus, were obtained simultaneously. The strategy is applicable to a wide range of sexually reproducing organisms, and could, therefore, accelerate the whole genome analysis of various organisms including fish, mollusks, amphibians, insects, plants, and even mammals.

Highlights

  • New-generation technologies enable whole-genome sequencing of any organism, assembling long, high-quality genomes from short reads generated by these technologies remains challenging

  • Construction of conventional linkage maps based on microsatellite markers generally is costly and time-consuming, and the maps are of low resolution and genomic coverage due to the limited number of markers, usually several hundreds to several thousands[5]

  • We initially speculated that whole-genome sequencing would be suitable to type SNPs in double haploid and/or haploid organisms because a single read would be sufficient

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Summary

Introduction

New-generation technologies enable whole-genome sequencing of any organism, assembling long, high-quality genomes from short reads generated by these technologies remains challenging. Several new methods to construct linkage maps and assemble contigs/scaffolds have been developed, including SNP array[6], RAD-seq[7], and genotyping-by-sequencing[8]. T. rubripes and T. stictonotus adults were sequenced to generate reference sequences with which to identify the source of each raw read in hybrid fry, and to call and map SNPs. Raw data were obtained from one round of sequencing on HiSeq 2500.

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