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URL: https://dx.doi.org/10.17504/protocols.io.betejeje

Authors: Kentaro Itokawa, Tsuyoshi Sekizuka, Masanori Hashino, Rina Tanaka, Makoto Kuroda

Group: Coronavirus Method Development Community

Summary: This protocol is folked from "ARTIC amplicon sequencing protocol for MinION for nCoV-2019" by Josh Quick to adapt it to illumina sequencers. Because the PCR products are fragmented and ligated with adapters, this protocol is not restricted to 250 PE mode of MiSeq. While the library preparation uses QiaSeq FX by Qiagen and is basically straight forward (as par kit instruction but set to 1/4 scale), some tweaks for much of simplicity and speed were added.Change from the previous version (V1): The amount of cDNA input to multiplex PCR is increased (now almost same amount to the ARTIC's original protocol) (Step 10). This change gives better results for samples with extremely low RNA copy. Amount of adapter solution input was corrected (Step 19).

Affiliations: National Institute of Infectious Diseases, Japan, National Institute of Infectious Diseases, Japan, National Institute of Infectious Diseases, Japan, National Institute of Infectious Diseases, Japan, National Institute of Infectious Diseases, Japan

Version: 2

Publication Date: 2020

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