title: Comparative performance of double-digest RAD sequencing across divergent arachnid lineages.
abstract: Next-generation sequencing technologies now allow researchers of non-model systems to perform genome-based studies without the requirement of a (often unavailable) closely related genomic reference. We evaluated the role of restriction endonuclease (RE) selection in double-digest restriction-site-associated DNA sequencing (ddRADseq) by generating reduced representation genome-wide data using four different RE combinations. Our expectation was that RE selections targeting longer, more complex restriction sites would recover fewer loci than RE with shorter, less complex sites. We sequenced a diverse sample of non-model arachnids, including five congeneric pairs of harvestmen (Opiliones) and four pairs of spiders (Araneae). Sample pairs consisted of either conspecifics or closely related congeneric taxa, and in total 26 sample pair analyses were tested. Sequence demultiplexing, read clustering and variant calling were performed in the pyRAD program. The 6-base pair cutter EcoRI combined with methylated site-specific 4-base pair cutter MspI produced, on average, the greatest numbers of intra-individual loci and shared loci per sample pair. As expected, the number of shared loci recovered for a sample pair covaried with the degree of genetic divergence, estimated with cytochrome oxidase I sequences, although this relationship was non-linear. Our comparative results will prove useful in guiding protocol selection for ddRADseq experiments on many arachnid taxa where reference genomes, even from closely related species, are unavailable.
authors.affiliation: Department of Biology, San Diego State University, 5500 Campanile Drive, San Diego, CA, 92182, USA.
authors.forename: Mercedes
authors.initials: M
authors.lastname: Burns
authors.affiliation: Department of Biology, San Diego State University, 5500 Campanile Drive, San Diego, CA, 92182, USA.
authors.forename: James
authors.initials: J
authors.lastname: Starrett
authors.affiliation: Department of Biology, San Diego State University, 5500 Campanile Drive, San Diego, CA, 92182, USA.|Department of Biology, University of California, 900 University Avenue, Riverside, CA, 92521, USA.
authors.forename: Shahan
authors.initials: S
authors.lastname: Derkarabetian
authors.affiliation: Department of Biology, San Diego State University, 5500 Campanile Drive, San Diego, CA, 92182, USA.|Department of Biology, University of California, 900 University Avenue, Riverside, CA, 92521, USA.
authors.forename: Casey H
authors.initials: CH
authors.lastname: Richart
authors.affiliation: Department of Biology, San Diego State University, 5500 Campanile Drive, San Diego, CA, 92182, USA.
authors.forename: Allan
authors.initials: A
authors.lastname: Cabrero
authors.affiliation: Department of Biology, San Diego State University, 5500 Campanile Drive, San Diego, CA, 92182, USA.
authors.forename: Marshal
authors.initials: M
authors.lastname: Hedin
mesh_terms: D000818:Animals; D001097:Arachnida / Q000145:classification*; D004262:DNA Restriction Enzymes; D016678:Genome; D059014:High-Throughput Nucleotide Sequencing; D017422:Sequence Analysis, DNA / Q000379:methods*
chemical_list: D004262:DNA Restriction Enzymes
keywords: Araneomorphae; Mygalomorphae; Opiliones; ddRADseq; genomics; non-model organism