10.5061/DRYAD.J897M
Shokralla, Shadi
University of Guelph
Porter, Teresita M.
McMaster University
Gibson, Joel F.
University of Guelph
Dobosz, Rafal
University of Guelph
Janzen, Daniel H.
University of Pennsylvania
Hallwachs, Winnie
University of Pennsylvania
Golding, G. Brian
McMaster University
Hajibabaei, Mehrdad
University of Guelph
Data from: Massively parallel multiplex DNA sequencing for specimen
identification using an Illumina MiSeq platform
Dryad
dataset
2016
Malaise trap
multiplexing tagging
Next Generation Sequencing
2016-02-18T00:00:00Z
2016-02-18T00:00:00Z
en
https://doi.org/10.1038/srep09687
6391608368 bytes
1
CC0 1.0 Universal (CC0 1.0) Public Domain Dedication
Genetic information is a valuable component of biosystematics, especially
specimen identification through the use of species-specific DNA barcodes.
Although many genomics applications have shifted to High-Throughput
Sequencing (HTS) or Next-Generation Sequencing (NGS) technologies, sample
identification (e.g., via DNA barcoding) is still most often done with
Sanger sequencing. Here, we present a scalable double dual-indexing
approach using an Illumina Miseq platform to sequence DNA barcode markers.
We achieved 97.3% success by using half of an Illumina Miseq flowcell to
obtain 658 base pairs of the cytochrome c oxidase I DNA barcode in 1,010
specimens from eleven orders of arthropods. Our approach recovers a
greater proportion of DNA barcode sequences from individuals than does
conventional Sanger sequencing, while at the same time reducing both per
specimen costs and labor time by nearly 80%. In addition, the use of HTS
allows the recovery of multiple sequences per specimen, for deeper
analysis of genetic variation in target gene regions.
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Area de Conservación Guanacaste
northwestern Costa Rica
Area de Conservación Guanacaste Costa Rica