Direct RNA Sequencing of the Coding Complete Influenza A Virus Genome.

ABSTRACT: For the first time, a coding complete genome of an RNA virus has been sequenced in its original form. Previously, RNA was sequenced by the chemical degradation of radiolabeled RNA, a difficult method that produced only short sequences. Instead, RNA has usually been sequenced indirectly by copying it into cDNA, which is often amplified to dsDNA by PCR and subsequently analyzed using a variety of DNA sequencing methods. We designed an adapter to short highly conserved termini of the influenza A virus genome to target the (-) sense RNA into a protein nanopore on the Oxford Nanopore MinION sequencing platform. Utilizing this method with total RNA extracted from the allantoic fluid of influenza rA/Puerto Rico/8/1934 (H1N1) virus infected chicken eggs (EID₅₀ 6.8 × 10⁹), we

SUBMITTER: Keller MW 

PROVIDER: S-EPMC6158192 | biostudies-literature | 2018 Sep

REPOSITORIES: biostudies-literature