Author: Monica Sentmanat; Evguenia Kouranova; Xiaoxia Cui
Title: One-step RNA extraction for RT-qPCR detection of 2019-nCoV Document date: 2020_4_5
ID: ihhu7nef_2
Snippet: We placed swabs directly in QE buffer. Prior to heat extraction, samples were vortexed and divided into two. One was used to create a positive control counterpart with 2019-nCoV plasmid DNA template added at 100 copies/ul. Although 2019-nCoV is an RNA virus, this positive DNA control ensures the assay is working while maximizing saftey and the need for Biosaftey Level 2 (BSL2) precautions, as is required for handling 2019-nCoV RNA material. An RN.....
Document: We placed swabs directly in QE buffer. Prior to heat extraction, samples were vortexed and divided into two. One was used to create a positive control counterpart with 2019-nCoV plasmid DNA template added at 100 copies/ul. Although 2019-nCoV is an RNA virus, this positive DNA control ensures the assay is working while maximizing saftey and the need for Biosaftey Level 2 (BSL2) precautions, as is required for handling 2019-nCoV RNA material. An RNA extraction control using HCT-116 human colorectal cancer cells at 10 cells/ul was included. Samples were then heated at 65°C for 15 min followed by 98°C for 2 min to inactivate proteinase K and then directly used for the test using a single probe and primer set (N1) as well as the set for RNaseP.
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