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_25
Snippet: 1. Vortex the sample again, take 50 ul of sample each to two PCR tubes marked with "+" and "-". 2. Add 5000 copies of positive RNA control of N transcript to the tube marked with "+". 3. (Extraction) Place both PCR tubes on a PCR machine and run the following program: 65°C, 15 min, 98°C, 2 min. 4. Take 2 ul each from "+" and "-" samples and assemble the one-step RT-qPCR reactions, containing 1.5 ul of each primer/probe set, 5 ul of 4x reaction .....
Document: 1. Vortex the sample again, take 50 ul of sample each to two PCR tubes marked with "+" and "-". 2. Add 5000 copies of positive RNA control of N transcript to the tube marked with "+". 3. (Extraction) Place both PCR tubes on a PCR machine and run the following program: 65°C, 15 min, 98°C, 2 min. 4. Take 2 ul each from "+" and "-" samples and assemble the one-step RT-qPCR reactions, containing 1.5 ul of each primer/probe set, 5 ul of 4x reaction mix and 11.5 ul of nuclease-free water. 5. Run the reactions on a QuantStudio™ 3 real-time PCR machine (ThermoFisher) using the following program:
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