WGS-SARS-CoV-2: Wastewater
After reverse transcription with 10 L RNA, a two-step PCR for target enrichment and sequence adapter with dual-barcode primer addition, was done. PCR products were purified by magnetic bead-based techniques followed by quantity and quality checks using the Qubit dsDNA high sensitivity assay kit (Life Technologies, USA) and gel-based electrophoreses, respectively. Libraries were equally pooled, and single strand circularized DNA was synthesized using the MGIEasy Dual Barcode Circularization kit. DNA nanoballs (DNBs) were constructed for both the ssDNA sample library and a dual barcoded balance library in parallel, which was then mixed 3:1 prior to sequencing on the DNBSEQ-G400 instrument at SAMRCs Genomics Centre