Why barcode? High-throughput multiplex sequencing of mitochondrial genomes for molecular systematics
Average rating
Cast your vote
You can rate an item by clicking the amount of stars they wish to award to
this item.
When enough users have cast their vote on this item, the average rating will also be shown.
Star rating
Your vote was cast
Thank you for your feedback
Thank you for your feedback
Issue date
2010-09-28Submitted date
2020-03-04Subject Terms
Gene sequencingPhylogenetics
DNA
Nucleotides
Bioinformatics
Metadata
Show full item recordAbstract
Mitochondrial 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.Citation
M. 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/gkq807Publisher
Oxford University Press (OUP)Journal
Nucleic Acids ResearchType
Journal ArticleItem Description
© 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.ISSN
0305-1048EISSN
1362-4962ae974a485f413a2113503eed53cd6c53
10.1093/nar/gkq807
Scopus Count