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Published on: June 3, 2019
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Scalable, Cost-Effective, and Decentralized DNA Barcoding with Oxford Nanopore Sequencing
Amrita Srivathsan1, Rudolf Meier2,3
1Center for Integrative Biodiversity Discovery, Leibniz Institute for Evolution and Biodiversity Science, Museum für Naturkunde, Berlin, Germany.
Methods in Molecular Biology (Clifton, N.J.)
|April 29, 2024
Summary
Oxford Nanopore
Area of Science:
- Genomics
- Biodiversity Research
- Molecular Biology
Background:
- DNA barcoding is crucial for biodiversity research but traditionally limited by costly and time-consuming Sanger sequencing.
- High-throughput sequencing technologies offer potential solutions for scalable DNA barcoding.
Purpose of the Study:
- To present a cost-effective laboratory workflow for DNA barcoding using MinION sequencing.
- To evaluate the efficiency and accuracy of MinION for large-scale species discovery and specimen identification.
Main Methods:
- Development of a laboratory workflow for tagged amplicon preparation, Oxford Nanopore Technologies (ONT) library preparation, and amplicon pool sequencing.
- Utilizing MinION and Flongle flow cells for sequencing.
- Analysis of MinION reads using the ONTbarcoder software.
Main Results:
- The MinION workflow provides highly accurate DNA barcodes, achieving 99.99% identity with Sanger sequencing results.
- Achieved a low cost per specimen (under $0.10 USD for MinION, $0.50 USD for Flongle).
- Demonstrated suitability for generating up to 10,000 barcodes per MinION flow cell.
Conclusions:
- MinION sequencing offers a cost-effective and efficient alternative to Sanger sequencing for DNA barcoding.
- This workflow supports large-scale species discovery and specimen identification.
- MinION is a viable option for researchers needing accessible DNA barcoding solutions.
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