Concentration and nucleic acid extraction of viruses from wastewater primary sludge v1

• DOI: 10.17504/protocols.io.36wgq3k1xlk5/v1
• Publication Date: 2024-03-02T17:22:42Z
• Authors: Ari N Machtinger; Olivia S Hershey; William J Bradshaw; Emily H Soice; Daniel P Rice; Michael R McLaren

This protocol details our workflow for performing concentration and total nucleic acid extraction from wastewater primary sludge (settled solids) the purposes of untargeted RNA DNA sequencing viruses present in wastewater. Protocol overview: In this protocol, 20 mL is concentrated to a final volume 400 uL using InnovaPrep Concentrating Pipette Select. Prior concentration, sample treated with Tween sonicated dissociate viral particles solids matrix. The then centrifuged remove larger solids. pellet discarded, supernatant filtered 0.45 um PES 75 mm filtration unit remaining suspended bacteria. filtrate Pipette, Ultra CPT tips recommended device settings modified processing protocol. Nucleic acids are extracted product Zymo quick-DNA/RNA Viral kit manufacturer few modifications that we have found be helpful.