ONTbarcoder and MinION barcodes aid biodiversity discovery and identification by everyone, for everyone

• DOI: 10.1186/s12915-021-01141-x
• Publication Date: 2021-09-29T08:20:39Z
• Authors: Amrita Srivathsan; Leshon Lee; Kazutaka Katoh; Emily Hartop; Sujatha Narayanan...; Johnathan Wong; Darren Yeo; Rudolf Meier
• Keywords: Minion; Barcode; OS X; Amplicon

DNA barcodes are a useful tool for discovering, understanding, and monitoring biodiversity which critical tasks at time of rapid loss. However, widespread adoption requires cost-effective simple barcoding methods. We here present workflow that satisfies these conditions. It was developed via "innovation through subtraction" thus minimal lab equipment, can be learned within days, reduces the barcode sequencing cost to < 10 cents, allows fast turnaround from specimen sequence by using portable MinION sequencer.We describe how tagged amplicons obtained sequenced with real-time sequencer in many settings (field stations, labs, citizen science schools). also provide amplicon coverage recommendations based on several runs latest generation flow cells ("R10.3") suggest each run generate > 10,000 specimens. Next, we novel software, ONTbarcoder, overcomes bioinformatics challenges posed reads. The software is compatible Windows 10, Macintosh, Linux, has graphical user interface (GUI), thousands standard laptop hours only two input files (FASTQ, demultiplexing file). document virtually identical Sanger Illumina same specimens (> 99.99%) evidence reads have improved rapidly since 2018.We propose way forward government agencies, universities, museums, schools because it combines low consumable capital scalability. Small projects use cell dongle ("Flongle") while large rely stopped re-used after collecting sufficient data given project.