title: WGS of Genome in a Bottle HG002 on Nanopore MinION with the R9.4 flow cell within the             NGS2 Study
design: Microbial gDNA was prepared for Nanopore sequencing using two                 library methods. For the Flongle flow cell runs, the direct ligation sequencing                 library kit (LSK-109 Oxford Nanopore UK) was used on individual bacteria and                 sequenced on dedicated flow cells. For the R9.4 flow cells runs, the individual                 bacteria strains as well as the 10 species mix was prepared using the LSK109 method                 with the native barcoding expansion kit (EXP-NBD104) and combined into one final                 library pool and sequenced together on a single flow cell. This ligation sequencing                 method is a non-PCR based library method that allows for direct sequencing of native                 DNA. Briefly, gDNA is reSINGLE using the NEBNext FFPE DNA Repair reagents (M6630,                 New England Biolabs Ipswich Ma) followed by dA-tailed using the NEBNext End                 Repair/dA-tailing module, and ligated to nanopore specific sequencing adapters.                 Sequencing was performed immediately after library synthesis.