title: Practical low-coverage genomewide sequencing of hundreds of individually barcoded samples for population and evolutionary genomics in nonmodel species.
abstract: Today most population genomic studies of nonmodel organisms either sequence a subset of the genome deeply in each individual or sequence pools of unlabelled individuals. With a step-by-step workflow, we illustrate how low-coverage whole-genome sequencing of hundreds of individually barcoded samples is now a practical alternative strategy for obtaining genomewide data on a population scale. We used a highly efficient protocol to generate high-quality libraries for ~6.5 USD from each of 876 Atlantic silversides (a teleost fish with a genome size ~730 Mb) that we sequenced to 1-4× genome coverage. In the absence of a reference genome, we developed a bioinformatic pipeline for mapping the genomic reads to a de novo assembled reference transcriptome. This provides an 'in silico' method for exome capture that avoids the complexities and expenses of using wet chemistry for target isolation. Using novel tools for analysis of low-coverage data, we extracted population allele frequencies, individual genotype likelihoods and polymorphism data for 2 504 335 SNPs across the exome for the 876 fish. To illustrate the use of the resulting data, we present a preliminary analysis of geographical patterns in the exome data and a comparison of complete mitochondrial genome sequences for each individual (constructed from the low-coverage data) that show population colonization patterns along the US east coast. With a total cost per sample of less than 50 USD (including sequencing) and ability to prepare 96 libraries in only 5 h, our approach adds a viable new option to the population genomics toolbox.
authors: Hopkins Marine Station, Department of Biology, Stanford University, 120 Oceanview Blvd., Pacific Grove, CA, 93950, USA.
authors: Nina Overgaard
authors: NO
authors: Therkildsen
authors: Hopkins Marine Station, Department of Biology, Stanford University, 120 Oceanview Blvd., Pacific Grove, CA, 93950, USA.
authors: Stephen R
authors: SR
authors: Palumbi
mesh_terms: D000818:Animals; D019295:Computational Biology / Q000379:methods*; D003365:Costs and Cost Analysis; D019143:Evolution, Molecular; D005399:Fishes / Q000145:classification* / Q000235:genetics*; D005787:Gene Frequency; D005828:Genetics, Population / Q000379:methods*; D023281:Genomics / Q000379:methods*; D058974:Phylogeography; D011110:Polymorphism, Genetic
keywords: exome capture; library preparation; low-coverage sequencing; mitogenomics; nonmodel population genomics