A method for determining the origin of crude drugs derived from animals using MinION, a compact next-generation sequencer

• Main Author: Nakanishi, Hiroaki
• Format: Other/Unknown Material
• Language: unknown
• Published: Zenodo 2022
• Subjects: Rangifer tarandus
• Online Access: https://doi.org/10.5061/dryad.w9ghx3frq

We evaluated whether MinION, an inexpensive, portable sequencer, can be applied for identifying the origin of crude drugs derived from animals . Standard and nonstandard crude drugs with different species of origin were examined. In addition, the standards mixed with nonstandard samples were used . As a target gene, cytochrome c oxidase I was amplified and sequenced. The Fast mode results had a slightly lower match ratio than High-accuracy mode, but the animals of origin were correctly determined by BLAST for all samples. For antler velvet derived from Rangifer tarandus , even the sequences were aligned based on Cervus elaphus , the animal of origin was determined correctly. Minor contents could be detected from mixtures of two animals, if the mixtures contained at least 19:1 mtDNA when the coverage allele-fraction threshold was 0.05. By contrast, in Fast mode, two sequences could not be separated due to the low accuracy of the base-calling in each read. For field work, the species of origin of crude drugs could be identified, by only simple DNA extraction and library preparation. Therefore, MinION appears to be a convenient tool for identifying the origin of crude drugs derived from animals. Funding provided by: Japan Society for the Promotion of Science Crossref Funder Registry ID: http://dx.doi.org/10.13039/501100001691 Award Number: JP19K07151