Shotgun Metagenomics of Hospital and Urban Wastewater in Dhaka, Bangladesh
Shotgun metagenomic libraries were constructed using the Illumina DNA Prep Kit (Illumina, San Diego, CA, USA) per the manufacturer's protocol. Briefly, genomic DNA was tagmented, followed by amplification with unique dual-index adapters and bead-based purification using the Enhanced PCR Mix supplied with the kit. Library fragment size distribution was confirmed using the LabChip GXII Touch 24 system (PerkinElmer, Waltham, MA, USA) and library concentrations were determined using the Qubit dsDNA High Sensitivity Assay Kit (Thermo Fisher Scientific, USA). Individual libraries were normalized to equimolar concentrations and pooled prior to sequencing on the Illumina NextSeq 500 platform (Illumina, USA) using a paired-end 2  150 bp configuration.