Rapid multiplex MinION nanopore sequencing workflow for Influenza A viruses.

ABSTRACT:

Background

Due to the frequent reassortment and zoonotic potential of influenza A viruses, rapid gain of sequence information is crucial. Alongside established next-generation sequencing protocols, the MinION sequencing device (Oxford Nanopore Technologies) has become a serious competitor for routine whole-genome sequencing. Here, we established a novel, rapid and high-throughput MinION multiplexing workflow based on a universal RT-PCR.

Methods

Twelve representative influenza A virus samples of multiple subtypes were universally amplified in a one-step RT-PCR and subsequently sequenced on the MinION instrument in conjunction with a barcoding library preparation kit from the rapid family and the MinIT performing live base-calling. The identical PCR products were sequenced on

SUBMITTER: King J 

PROVIDER: S-EPMC7468549 | biostudies-literature | 2020 Sep

REPOSITORIES: biostudies-literature