Tissue Culture (tc) PBS
Jacquelina.Woods
Published: 2021-12-02 DOI: 10.17504/protocols.io.byajpscn
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
1X PBS
1.
Dissolve components in deionized or ultrapure water to 1L.
1.1.
8.0g
1.2.
0.2g
1.3.
0.12g
1.4.
0.91g
2.
Adjust 7.5
3.
Autoclave 121°C``0h 15m 0s
4. For 2X tc PBS, double components in 1.1 - 1.4
Store at 2-8°C
Note
10X tc PBS
5. Alternatively, 1X and 2X PBS can be made by dilution of 10X PBS into sterile deionized or ultrapure water.
Note
