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Real-time DNA barcoding in a remote rainforest using nanopore sequencing

Aaron Pomerantz, Nicolás Peñafiel, Alejandro Arteaga, Lucas Bustamante, Frank Pichardo, Luis A. Coloma, César L. Barrio-Amorós, David Salazar-Valenzuela, Stefan Prost
doi: https://doi.org/10.1101/189159
Aaron Pomerantz
1Department of Integrative Biology, University of California, Berkeley, CA, USA
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  • For correspondence: Pomerantz_aaron@berkeley.edu stprost@stanford.edu
Nicolás Peñafiel
2Centro de Investigación de la Biodiversidad y Cambio Climático (BioCamb) e Ingeniería en Biodiversidad y Recursos Genéticos, Facultad de Ciencias de Medio Ambiente, Universidad Tecnológica Indoamérica, Machala y Sabanilla, Quito, Ecuador
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Alejandro Arteaga
3Tropical Herping, Quito, Ecuador
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Lucas Bustamante
3Tropical Herping, Quito, Ecuador
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Frank Pichardo
3Tropical Herping, Quito, Ecuador
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Luis A. Coloma
4Centro Jambatu de Investigación y Conservación de Anfibios, Fundación Otonga, Quito, Ecuador
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César L. Barrio-Amorós
5Doc Frog Expeditions, Uvita, Costa Rica
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David Salazar-Valenzuela
2Centro de Investigación de la Biodiversidad y Cambio Climático (BioCamb) e Ingeniería en Biodiversidad y Recursos Genéticos, Facultad de Ciencias de Medio Ambiente, Universidad Tecnológica Indoamérica, Machala y Sabanilla, Quito, Ecuador
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Stefan Prost
1Department of Integrative Biology, University of California, Berkeley, CA, USA
6Program for Conservation Genomics, Department of Biology, Stanford University, Stanford, CA, USA
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  • For correspondence: Pomerantz_aaron@berkeley.edu stprost@stanford.edu
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Abstract

Advancements in portable scientific instruments provide promising avenues to expedite field work in order to understand the diverse array of organisms that inhabit our planet. Here we tested the feasibility for in situ molecular analyses of endemic fauna using a portable laboratory fitting within a single backpack, in one of the world’s most imperiled biodiversity hotspots: the Ecuadorian Chocó rainforest. We utilized portable equipment, including the MinION DNA sequencer (Oxford Nanopore Technologies) and miniPCR (miniPCR), to perform DNA extraction, PCR amplification and real-time DNA barcode sequencing of reptile specimens in the field. We demonstrate that nanopore sequencing can be implemented in a remote tropical forest to quickly and accurately identify species using DNA barcoding, as we generated consensus sequences for species resolution with an accuracy of >99% in less than 24 hours after collecting specimens. In addition, we generated sequence information at Universidad Tecnológica Indoamérica in Quito for the recently re-discovered Jambato toad Atelopus ignescens, which was thought to be extinct for 28 years, a rare species of blind snake Trilepida guayaquilensis, and two undescribed species of Dipsas snakes. In this study we establish how mobile laboratories and nanopore sequencing can help to accelerate species identification in remote areas (especially for species that are difficult to diagnose based on characters of external morphology), be applied to local research facilities in developing countries, and rapidly generate information for species that are rare, endangered and undescribed, which can potentially aid in conservation efforts.

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The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license.
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Posted September 15, 2017.
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Real-time DNA barcoding in a remote rainforest using nanopore sequencing
Aaron Pomerantz, Nicolás Peñafiel, Alejandro Arteaga, Lucas Bustamante, Frank Pichardo, Luis A. Coloma, César L. Barrio-Amorós, David Salazar-Valenzuela, Stefan Prost
bioRxiv 189159; doi: https://doi.org/10.1101/189159
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Real-time DNA barcoding in a remote rainforest using nanopore sequencing
Aaron Pomerantz, Nicolás Peñafiel, Alejandro Arteaga, Lucas Bustamante, Frank Pichardo, Luis A. Coloma, César L. Barrio-Amorós, David Salazar-Valenzuela, Stefan Prost
bioRxiv 189159; doi: https://doi.org/10.1101/189159

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