Dec 02, 2021

Public workspacePrimer TE (10mM Tris, 0.1mM EDTA, pH8.0)

DOI
dx.doi.org/10.17504/protocols.io.byccpssw
  • 1FDA
  • GenomeTrakr
    Tech. support email: genomeTrakr@fda.hhs.gov
Jessica L Jones
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DOI: dx.doi.org/10.17504/protocols.io.byccpssw
Protocol Citation: Jacquelina.Woods 2021. Primer TE (10mM Tris, 0.1mM EDTA, pH8.0). protocols.io https://dx.doi.org/10.17504/protocols.io.byccpssw
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 17, 2021
Last Modified: December 02, 2021
Protocol Integer ID: 53348
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
ReagentTris (1 M) pH 8.0 RNase-freeThermo Fisher ScientificCatalog #AM9856
ReagentEDTA (0.5 M) pH 8.0 RNase-freeThermo Fisher ScientificCatalog #AM9261
ReagentNuclease-Free WaterThermo Fisher ScientificCatalog #AM9937

Protocol materials
ReagentTris (1 M) pH 8.0 RNase-freeThermo Fisher ScientificCatalog #AM9856
ReagentEDTA (0.5 M) pH 8.0 RNase-freeThermo Fisher ScientificCatalog #AM9261
ReagentNuclease-Free WaterThermo Fisher ScientificCatalog #AM9937
ReagentTris (1 M) pH 8.0 RNase-freeThermo Fisher ScientificCatalog #AM9856
ReagentEDTA (0.5 M) pH 8.0 RNase-freeThermo Fisher ScientificCatalog #AM9261
ReagentNuclease-Free WaterThermo Fisher ScientificCatalog #AM9937
Mix components together.
Amount100 µL ReagentTris (1 M) pH 8.0 RNase-freeThermo Fisher ScientificCatalog #AM9856,, or equivalent.

Amount20 µL ReagentEDTA (0.5 M) pH 8.0 RNase-freeThermo Fisher ScientificCatalog #AM9261,, or equivalent.

Amount9.88 mL ReagentNuclease-Free WaterThermo Fisher ScientificCatalog #AM9937 , or equivalent.

Store at TemperatureRoom temperature .