Protocol Citation: Jacquelina.Woods 2021. 6M GITC. protocols.io https://dx.doi.org/10.17504/protocols.io.bybbpsin
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 16, 2021
Last Modified: December 02, 2021
Protocol Integer ID: 53315
Keywords: rna extraction, extraction of rna, viral concentrate, applied nutrition for genometrakr, wastewater, wastewater collection, rna, genometrakr, pandemic response project, genome sequencing, gitc this method, applied nutrition, viral, center for food safety, data submission to ncbi, extraction, monitoring sar, gitc
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 extraction of RNA from viral concentrates using the RNeasy and Zymo kits.
Materials
Guanidine IsothiocyanateThermo FisherCatalog #15535016
Protocol materials
Guanidine IsothiocyanateThermo FisherCatalog #15535016
Troubleshooting
Aseptically mix 7.09 g Guanidine IsothiocyanateThermo FisherCatalog #15535016 in sterile distilled or ultrapure water to 4.5 mL
Bring total volume up to 10 mL with additional sterile distilled or ultrapure water
Store at Room temperature in the dark (or in light occluding tube)
Note
Solution is only stable for 1 month after preparation.