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URL: https://dx.doi.org/10.17504/protocols.io.bi6skhee

Authors: Stephanie Loeb, Katy Graham, Marlene Wolfe, Krista Wigginton, Alexandria Boehm

Group: Coronavirus Method Development Community, Wastewater-based epidemiology working group

Summary: OverviewThis SOP describes pre-analytical procedures to be followed for the isolation and identification of SARS-CoV-2 RNA in primary settled solids samples from wastewater treatment plants. This protocal should be paired with our digital PCR protocal. The protocol follows the approximate workflow:Separate and aliquot sample (~4 hrs)Spike with BCoV (~1 hr)Extract RNA (~ 12 hours total, may be split into 2-3 steps/days)Inhibitor removal (~1 hr)Biosafety ConcernsConcentration and extraction procedures that utilize raw samples must adhere to strict Biosafety Level 2+ procedures. These procedures should be performed in a dedicated room. Downstream products may be handled using standard laboratory safety guidelines. This protocol was developed by Stephanie Loeb, Katy Graham, Marlene Wolfe, Krista Wigginton, and Alexandria Boehm at Stanford University and University of Michigan.

Affiliations: Stanford University, Stanford University, Stanford University, University of Michigan, Stanford University

Version: 1

Publication Date: 2020

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