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. 2021 May 6;13(5):847. doi: 10.3390/v13050847

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© 2021 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).

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Workflow overview of the amplicon-based nanopore sequencing for Hantaan virus (HTNV) used in this study. Total RNA was extracted from rodent lung tissues, and cDNA was synthesized from 1 µg of total RNA with a random hexamer and OSM55. cDNA was amplified using HTNV-specific primer mixtures for library preparation. Barcoded libraries were pooled, ligated to sequencing adapter, and sequenced using the MinION device with FLO-MIN106 (R9.4) flow cell for 8 h. The basecalled sequences were combined into a single FASTA file, and the reads were mapped to the reference genome sequences. The consensus sequences were extracted for downstream analysis.