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Title
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Using next-generation sequencing to improve DNA barcoding: lessons from a small-scale study of wild bee species (Hymenoptera, Halictidae)
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Author
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Abstract
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| The parallel sequencing of targeted amplicons is a scalable application of next-generation sequencing (NGS) that can advantageously replace Sanger sequencing in certain DNA barcoding studies. It can be used to sequence different PCR products simultaneously, including co-amplified products. Here, we explore this approach by simultaneously sequencing five markers (including the DNA barcode and a diagnostic marker of Wolbachia) in 12 species of Halictidae that were previously DNA barcoded using Sanger sequencing. Consensus sequences were obtained from fresh bees with success rates of 74-100% depending on the DNA fragment. They improved the phylogeny of the group, detected Wolbachia infections (in 8/21 specimens) and characterised haplotype variants. Sequencing cost per marker and per specimen (11.43 a,notsign) was estimated to decrease (< 5.00 a,notsign) in studies aiming for a higher throughput. We provide guidelines for selecting NGS or Sanger sequencing depending on the goals of future studies. |
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Language
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English
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Source (journal)
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Apidologie. - Versailles
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Publication
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Versailles
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2018
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ISSN
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0044-8435
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DOI
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10.1007/S13592-018-0594-Y
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Volume/pages
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49
:5
(2018)
, p. 671-685
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ISI
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000451113900014
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Full text (Publisher's DOI)
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Full text (publisher's version - intranet only)
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