Jul 28, 2020
RT-LAMP SARS-CoV-2 positive control stock
This protocol is a draft, published without a DOI.
- Christina Newman1,
- Andrea Weiler1,
- Dawn Dudley1,
- Mitchell Ramuta1,
- Amelia Haj1,
- Shelby O'Connor1,
- David O'Connor1
- 1University of Wisconsin-Madison
- Coronavirus Method Development Community
Protocol Citation: Christina Newman, Andrea Weiler, Dawn Dudley, Mitchell Ramuta, Amelia Haj, Shelby O'Connor, David O'Connor 2020. RT-LAMP SARS-CoV-2 positive control stock. protocols.io https://protocols.io/view/rt-lamp-sars-cov-2-positive-control-stock-biz8kf9w
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
We use this protocol and it's working
Created: July 25, 2020
Last Modified: August 02, 2020
Protocol Integer ID: 39712
Materials
MATERIALS
1X PBS (Phosphate-buffered saline )
dH2O
1.5 ml tubes
PCR strip tubes
Heat-inactivated SARS-CoV-2ATCCCatalog #ATCC® VR-1986HK™
Using either the company-provided stock concentration or a qRT-PCR determined stock concentration, dilute virus in dH2O to create a 1e9 copies/ml stock virus.
Create a 1e6 copies/ml stock virus by diluting 1e9 copies/ml stock 1:1000 in 50% dH2O and 50% 1x PBS solution.
Create a ready-to-use positive control stock (1e3 copies per 3 µl) for RT-LAMP saliva testing assay by diluting 100 µl of the 1e6 copies/ml stock in 200 µl of 50% dH2O/50% 1x PBS solution.
Prepare 10 µl aliquots of ready-to-use positive control stock in PCR strip tubes.