2022-W34 Stockholm Amplified SARS-CoV-2 RNA from wastewater
wastewater metagenome
Untreated wastewater was collected in Stockholm at Käppalaverket, Bromma reningsverk and Henriksdal reningsverk on 2022-08-22 over 24 hours using flow compensated samplers. The samples were then combined flow-proportionally into a composite sample for the city. Viral genomic material was concentrated and extracted by the direct capture method using the Maxwell RSC Enviro Total Nucleic Acid Kit (Promega Corporation, Madison, WI, USA, Cat # AS1831) according to the manufacturer's protocol (Promega, # TM663 Rev 4/22) at SEEC-KTH. Sequencing libraries were made using Ion AmpliSeq SARS-CoV-2 Insight Research Assay (Thermo Fisher Scientific, Waltham, MA, USA, Cat # A51306) and the SuperScript™ VILO™ cDNA Synthesis Kit (Thermo Fisher, # 11754050) according to the manufacturer's protocols “Reverse transcribe RNA with the SuperScript™ VILO™ cDNA Synthesis Kit” (Thermo Fisher, # MAN0019277, Rev B.0, pp. 3-4) and “Prepare libraries on the Ion Chef™ Instrument” (Thermo Fisher, # MAN0024915 Rev B.0, Ch 4, pp. 42-43) at the National Genomic Infrastructure (NGI) at SciLifeLab in Uppsala, Sweden. Sequencing was performed on the Ion S5 XL System (Thermo Fisher, # A27214) multiplexing using Torrent Suite Software version 5.16.1 configured according to the manufacturer's protocol (Thermo Fisher, # MAN0024915 Rev B.0, Ch 5) at NGI.