Data from: A sequel to Sanger: amplicon sequencing that scales

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Description:

Although high-throughput sequencers (HTS) have largely displaced their Sanger counterparts, the short read lengths and high error rates of most platforms constrain their utility for amplicon sequencing. The present study tests the capacity of single molecule, real-time (SMRT) sequencing implemented on the SEQUEL platform to overcome these limitations, employing 658 bp amplicons of the mitochondrial cytochrome c oxidase I gene as a model system. By examining templates from more than 5,000 species and 20,000 specimens, the performance of SMRT sequencing was tested with amplicons showing wide variation in GC composition and varied sequence attributes. SMRT and Sanger sequences were very similar, but SMRT sequencing provided more complete coverage, especially for amplicons with homopolymer tracts. Because it can characterize amplicon pools from 10,000 DNA extracts in a single run, the SEQUEL reduces costs 40-fold from Sanger analysis. Reflecting the capacity of each instrument to recover sequences from more than five million DNA extracts a year, this platform facilitates massive amplicon characterization.

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Auteur(s) :

Hebert, Paul D.N.

Hebert, Paul D.N.

University of Guelph

, Braukmann, Thomas W.A.

Braukmann, Thomas W.A.

University of Guelph

, Prosser, Sean W.J.

Prosser, Sean W.J.

University of Guelph

, Ratnasingham, Sujeevan

Ratnasingham, Sujeevan

University of Guelph

, deWaard, Jeremy R.

deWaard, Jeremy R.

University of Guelph

, Ivanova, Natalia V.

Ivanova, Natalia V.

University of Guelph

, Janzen, Daniel H.

Janzen, Daniel H.

University of Pennsylvania

, Hallwachs, Winnie

Hallwachs, Winnie

University of Pennsylvania

, Naik, Suresh

Naik, Suresh

University of Guelph

, Sones, Jayme E.

Sones, Jayme E.

University of Guelph

et Zakharov, Evgeny V.

Zakharov, Evgeny V.

University of Guelph

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Dépôt source:

Dryad

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License:

https://creativecommons.org/publicdomain/zero/1.0/

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URL:

https://doi.org/10.5061/dryad.4368h

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Date de publication:

2019-01-12

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Mots-clés (en):

nucleotide composition, SMRT sequencing, et homopolymer

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Citation selon les normes APA:

Data from: A sequel to Sanger: amplicon sequencing that scales. (2019). [Data set]. Dryad. https://doi.org/10.5061/dryad.4368h

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