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Recovery of DNA barcodes from blackfly museum specimens ( D iptera: S imuliidae) using primer sets that target a variety of sequence lengths
Author(s) -
HernándezTriana L. M.,
Prosser S. W.,
RodríguezPerez M. A.,
Chaverri L. G.,
Hebert P. D. N.,
Ryan Gregory T.
Publication year - 2014
Publication title -
molecular ecology resources
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.96
H-Index - 136
eISSN - 1755-0998
pISSN - 1755-098X
DOI - 10.1111/1755-0998.12208
Subject(s) - biology , primer (cosmetics) , phylogenetic tree , dna , dna sequencing , genetics , gene , computational biology , chemistry , organic chemistry
In this study, we evaluated the efficacy of various primers for the purpose of DNA barcoding old, pinned museum specimens of blackflies ( D iptera: S imuliidae). 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.