title: Recovery of DNA barcodes from blackfly museum specimens (Diptera: Simuliidae) using primer sets that target a variety of sequence lengths.
abstract: In this study, we evaluated the efficacy of various primers for the purpose of DNA barcoding old, pinned museum specimens of blackflies (Diptera: Simuliidae). We analysed 271 pinned specimens representing two genera and at least 36 species. Due to the age of our material, we targeted overlapping DNA fragments ranging in size from 94 to 407 bp. We were able to recover valid sequences from 215 specimens, of which 18% had 500- to 658-bp barcodes, 36% had 201- to 499-bp barcodes and 46% had 65- to 200-bp barcodes. Our study demonstrates the importance of choosing suitable primers when dealing with older specimens and shows that even very short sequences can be diagnostically informative provided that an appropriate gene region is used. Our study also highlights the lack of knowledge surrounding blackfly taxonomy, and we briefly discuss the need for further phylogenetic studies in this socioeconomically important family of insects.
authors.affiliation: Biodiversity Institute of Ontario, University of Guelph, Guelph, Ontario, N1G 2W1, Canada.
authors.forename: L M
authors.initials: LM
authors.lastname: Hernández-Triana
authors.forename: S W
authors.initials: SW
authors.lastname: Prosser
authors.forename: M A
authors.initials: MA
authors.lastname: Rodríguez-Perez
authors.forename: L G
authors.initials: LG
authors.lastname: Chaverri
authors.forename: P D N
authors.initials: PD
authors.lastname: Hebert
authors.forename: T Ryan
authors.initials: TR
authors.lastname: Gregory
mesh_terms: D000818:Animals; D058893:DNA Barcoding, Taxonomic / Q000379:methods*; D017931:DNA Primers / Q000235:genetics*; D003576:Electron Transport Complex IV / Q000235:genetics; D019476:Insect Proteins / Q000235:genetics; D008969:Molecular Sequence Data; D009144:Museums; D010802:Phylogeny; D012843:Simuliidae / Q000145:classification* / Q000201:enzymology / Q000235:genetics*
chemical_list: D017931:DNA Primers; D019476:Insect Proteins; D003576:Electron Transport Complex IV
keywords: COI; DNA barcoding; Simuliidae; blackflies; museum specimens