title: metagenome of hospital wastewater:untreated
design: Long-read DNA libraries for nanopore sequencing were prepared with the use of the gDNA ligation sequencing technology Native Barcoding Kit 24 V14 (SQK-NBD114.24; Oxford Nanopore Technologies), according to the manufacturers protocol. Four samples containing 500 ng of gDNA each were multiplexed in each of the three sequencing runs. In the first step, DNA was fragmented and end-repaired using the NEBNextCompanion Module for Oxford Nanopore Ligation Sequencing (New England Biolabs). Next, a unique barcode was ligated to each sample; the samples were pooled, and sequencing adapters were ligated. The libraries were quantified with the Qubit dsDNA HS Assay Kit (ThermoFisher) and sequenced using the MinION MK1C portable device (ONT) and FLO-MIN 112 Flow Cells R.10.4 (ONT).