Comprehensive Wastewater Sequencing Reveals Community and Variant Dynamics of the Collective Human Virome
DNA and RNA were extracted from wastewater samples using the Qiagen QIAamp VIRAL RNA Mini Kit. RNA extracts were converted to cDNA and the cDNA and DNA mix was used for library construction using Twist Library Preparation EF 2.0 Kit and Twist Universal Adaptor System (Twist Biosciences). The libraries were pooled, a maximum of 16 samples per pool, followed by hybridization using the Twist Comprehensive Viral Research Panel (Twist Biosciences). The post-capture pool was further PCR amplified for 12 cycles and final libraries were sequenced on Illumina NovaSeq 6000 SP flow cell, to generate 2150 bp paired-end reads.