6 million sequences can be downloaded as “public data” (April 2021: ). 8 million animal barcodes are available for searches in the database “BOLD Systems” of which ca. After 15 years and the investment of > 200 million USD in facilities and barcoding, ca. Obtaining barcodes was too complicated and expensive despite the development of effective DNA extraction protocols, fast Sanger sequencing protocols, and the establishment of the Canadian Centre for DNA Barcoding (CCDB), which is the primary sequencing facility for the International Barcode of Life Consortium (iBOL: ). Small projects can use the flow cell dongle (“Flongle”) while large projects can rely on MinION flow cells that can be stopped and re-used after collecting sufficient data for a given project.ĭNA sequences have been used for identification and taxonomic purposes for decades, but for most of this time, they have been akin to mobile phones in the 1990s: of limited value due to sparse signal coverage and high cost. We propose that barcoding with MinION is the way forward for government agencies, universities, museums, and schools because it combines low consumable and capital cost with scalability. We document that MinION barcodes are virtually identical to Sanger and Illumina barcodes for the same specimens (> 99.99%) and provide evidence that MinION flow cells and reads have improved rapidly since 2018. The software is compatible with Windows 10, Macintosh, and Linux, has a graphical user interface (GUI), and can generate thousands of barcodes on a standard laptop within hours based on only two input files (FASTQ, demultiplexing file). Next, we present a novel software, ONTbarcoder, which overcomes the bioinformatics challenges posed by MinION reads. It was developed via “innovation through subtraction” and thus requires minimal lab equipment, can be learned within days, reduces the barcode sequencing cost to 10,000 specimens. We here present a workflow that satisfies these conditions. However, widespread adoption of barcodes requires cost-effective and simple barcoding methods. DNA barcodes are a useful tool for discovering, understanding, and monitoring biodiversity which are critical tasks at a time of rapid biodiversity loss.
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