Protocol Citation: Jacquelina.Woods 2021. 5N HCl. protocols.io https://dx.doi.org/10.17504/protocols.io.byfsptne
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
Created: September 22, 2021
Last Modified: December 02, 2021
Protocol Integer ID: 53458
Keywords: rapid concentration of intact virus, 5n hcl, wastewater, pandemic response project, applied nutrition for genometrakr, intact virus, wastewater collection, rna extraction, center for food safety, genometrakr, pandemic, genome sequencing, qpcr detection
Abstract
This method was developed at the FDA’s Center for Food Safety and Applied Nutrition for GenomeTrakr’s pandemic response project, monitoring SARS-CoV-2 variants in wastewater. Protocols developed for this project cover wastewater collection, concentration, RNA extraction, RT-qPCR detection, library prep, genome sequencing, quality control checks, and data submission to NCBI. This method provides a reagent formula required in the rapid concentration of intact viruses from wastewater using a combination of PEG precipitation and ultracentrifugation.
Materials
Hydrochloric Acid (HCl)Fisher ScientificCatalog #6000710
Deionized or ultrapure water
Sterile container (e.g., 50 ml conical tube, Fisher Scientific Cat # 14-432-22, or equivalent)
Protocol materials
Hydrochloric Acid (HCl)Fisher ScientificCatalog #6000710
Troubleshooting
Measure 14.6 mL of deionized or ultrapure water into sterile container.
Slowly add 10.4 mL Hydrochloric Acid (HCl)Fisher ScientificCatalog #6000710 , or equivalent to the water.
Safety information
This solution should be prepared under a chemical fume hood.
Store at Room temperature .