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dc.contributor.authorTimmermans, MJTN
dc.contributor.authorDodsworth, S
dc.contributor.authorCulverwell, CL
dc.contributor.authorBocak, L
dc.contributor.authorAhrens, D
dc.contributor.authorLittlewood, T
dc.contributor.authorPons, J
dc.contributor.authorVogler, AP
dc.date.accessioned2020-03-30T09:01:54Z
dc.date.available2020-03-30T09:01:54Z
dc.date.issued2010-09-28
dc.date.submitted2020-03-04
dc.identifier.citationM. J. T. N. Timmermans, S. Dodsworth, C. L. Culverwell, L. Bocak, D. Ahrens, D. T. J. Littlewood, J. Pons, A. P. Vogler, Why barcode? High-throughput multiplex sequencing of mitochondrial genomes for molecular systematics, Nucleic Acids Research, Volume 38, Issue 21, 1 November 2010, Page e197, https://doi.org/10.1093/nar/gkq807en_US
dc.identifier.issn0305-1048
dc.identifier.doi10.1093/nar/gkq807
dc.identifier.urihttp://hdl.handle.net/10141/622664
dc.description.abstractMitochondrial genome sequences are important markers for phylogenetics but taxon sampling remains sporadic because of the great effort and cost required to acquire full-length sequences. Here, we demonstrate a simple, cost-effective way to sequence the full complement of protein coding mitochondrial genes from pooled samples using the 454/Roche platform. Multiplexing was achieved without the need for expensive indexing tags (‘barcodes’). The method was trialled with a set of long-range polymerase chain reaction (PCR) fragments from 30 species of Coleoptera (beetles) sequenced in a 1/16th sector of a sequencing plate. Long contigs were produced from the pooled sequences with sequencing depths ranging from ∼10 to 100× per contig. Species identity of individual contigs was established via three ‘bait’ sequences matching disparate parts of the mitochondrial genome obtained by conventional PCR and Sanger sequencing. This proved that assembly of contigs from the sequencing pool was correct. Our study produced sequences for 21 nearly complete and seven partial sets of protein coding mitochondrial genes. Combined with existing sequences for 25 taxa, an improved estimate of basal relationships in Coleoptera was obtained. The procedure could be employed routinely for mitochondrial genome sequencing at the species level, to provide improved species ‘barcodes’ that currently use the cox1 gene only.en_US
dc.language.isoenen_US
dc.publisherOxford University Press (OUP)en_US
dc.rightsopenAccessen_US
dc.rights.urihttp://creativecommons.org/licenses/by-nc/2.5
dc.titleWhy barcode? High-throughput multiplex sequencing of mitochondrial genomes for molecular systematicsen_US
dc.typeJournal Articleen_US
dc.identifier.eissn1362-4962
dc.identifier.journalNucleic Acids Researchen_US
dc.identifier.volume38en_US
dc.identifier.issue21en_US
dc.identifier.startpagee197 - e197en_US
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dc.embargoNot knownen_US
elements.import.authorTimmermans, MJTNen_US
elements.import.authorDodsworth, Sen_US
elements.import.authorCulverwell, CLen_US
elements.import.authorBocak, Len_US
elements.import.authorAhrens, Den_US
elements.import.authorLittlewood, DTJen_US
elements.import.authorPons, Jen_US
elements.import.authorVogler, APen_US
dc.description.nhm© The Author(s) 2010. Published by Oxford University Press. This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.5), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited. The attached file is the published pdf.en_US
dc.subject.nhmGene sequencingen_US
dc.subject.nhmPhylogeneticsen_US
dc.subject.nhmDNAen_US
dc.subject.nhmNucleotidesen_US
dc.subject.nhmBioinformaticsen_US
refterms.dateFOA2020-03-30T09:01:55Z


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