2X PEG

Jacquelina.Woods

Published: 2021-12-02 DOI: 10.17504/protocols.io.bycgpstw

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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.

Steps

1.

Dissolve components in ~700mL deionized or ultrapure water. Heating to 50°C is recommended to facilitate dissolution.

1.1.

200g , or equivalent

1.2.

34.8g, or equivalent.

2.

Bring total volume up to 1Lwith deionized or ultrapure water.

3.

Autoclave 121°C``0h 15m 0s.

4.

Store at 2-8Room temperatureprotected from light.

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