Author: Devon Chandler-Brown; Anna M. Bueno; Oguzhan Atay; David S. Tsao
Title: A Highly Scalable and Rapidly Deployable RNA Extraction-Free COVID-19 Assay by Quantitative Sanger Sequencing Document date: 2020_4_10
ID: eui41zyg_34
Snippet: qPCR were performed using the N1 primers and TaqMan probes provided in the 2019-nCoV RUO Kit manufactured by IDT (cat. # 10006605). Amplification was performed as described in the CDC EUA protocol. Briefly, 2 µL of synthetic RNA template diluted in RNAse-free TE + 0.05% Tween-20 to 5, 50, 500, or 2500 GCE/µL were added to each reaction.. RNA samples were combined with water, TaqPath 1-Step RT-qPCR Master Mix, primers (1.5 µL to a final concent.....
Document: qPCR were performed using the N1 primers and TaqMan probes provided in the 2019-nCoV RUO Kit manufactured by IDT (cat. # 10006605). Amplification was performed as described in the CDC EUA protocol. Briefly, 2 µL of synthetic RNA template diluted in RNAse-free TE + 0.05% Tween-20 to 5, 50, 500, or 2500 GCE/µL were added to each reaction.. RNA samples were combined with water, TaqPath 1-Step RT-qPCR Master Mix, primers (1.5 µL to a final concentration of 500 nM), and probes to a total final volume of 20 µL. The reaction mixture was amplified and probe fluorescence was detected using a Mastercycler ep realplex Real-time PCR System. The first cycle above threshold was estimated (Ct) was performed with default settings using realplex software.
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