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    Using next-generation sequencing to improve DNA barcoding: lessons from a small-scale study of wild bee species (Hymenoptera, Halictidae)

    Authors
    Sonet, G.
    Pauly, A.
    Nagy, ZT.
    Virgilio, M.
    Jordaens, K.
    Van Houdt, J.
    Worms, S.
    De Meyer, M.
    Backeljau, T.
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    Discipline
    Biological sciences
    Subject
    Invertebrates
    Audience
    Scientific
    Date
    2018
    Metadata
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    Description
    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 ) was estimated to decrease (< 5.00 ) in studies aiming for a higher throughput. We provide guidelines for selecting NGS or Sanger sequencing depending on the goals of future studies.
    Citation
    Sonet, G.; Pauly, A.; Nagy, ZT.; Virgilio, M.; Jordaens, K.; Van Houdt, J.; Worms, S.; De Meyer, M.; Backeljau, T. (2018). Using next-generation sequencing to improve DNA barcoding: lessons from a small-scale study of wild bee species (Hymenoptera, Halictidae). , Apidologie, DOI: https://doi.org/10.1007/s13592-018-0594-y.
    Identifiers
    uri: https://orfeo.belnet.be/handle/internal/12057
    doi: https://doi.org/10.1007/s13592-018-0594-y
    url: https://link.springer.com/article/10.1007/s13592-018-0594-y#citeas
    Type
    Article
    Peer-Review
    Yes
    Language
    eng
    Links
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