Multiplex PCR method for MinION and Illumina sequencing of Zika and other virus genomes directly from clinical samples.

ABSTRACT: Genome sequencing has become a powerful tool for studying emerging infectious diseases; however, genome sequencing directly from clinical samples (i.e., without isolation and culture) remains challenging for viruses such as Zika, for which metagenomic sequencing methods may generate insufficient numbers of viral reads. Here we present a protocol for generating coding-sequence-complete genomes, comprising an online primer design tool, a novel multiplex PCR enrichment protocol, optimized library preparation methods for the portable MinION sequencer (Oxford Nanopore Technologies) and the Illumina range of instruments, and a bioinformatics pipeline for generating consensus sequences. The MinION protocol does not require an Internet connection for analysis, making it suitable for field applicat

SUBMITTER: Quick J 

PROVIDER: S-EPMC5902022 | biostudies-literature | 2017 Jun

REPOSITORIES: biostudies-literature