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_6
Snippet: To determine the limit of detection of qSanger COVID-19, we performed assays on dilutions of SARS-COV-2 RNA corresponding to 0 GCE, 10 GCE, 100 GCE, 1000 GCE, or 5000 GCE. Four replicates at each dilution were assayed by both qSanger and qPCR. As expected, all 4 replicates of 0 GCE were negative for COVID-19 by qPCR, and addition of 10 or more SARS-CoV-2 GCE exhibited a clear logarithmic decrease in qPCR cycle threshold (Ct) (Fig. 2B) . Similarl.....
Document: To determine the limit of detection of qSanger COVID-19, we performed assays on dilutions of SARS-COV-2 RNA corresponding to 0 GCE, 10 GCE, 100 GCE, 1000 GCE, or 5000 GCE. Four replicates at each dilution were assayed by both qSanger and qPCR. As expected, all 4 replicates of 0 GCE were negative for COVID-19 by qPCR, and addition of 10 or more SARS-CoV-2 GCE exhibited a clear logarithmic decrease in qPCR cycle threshold (Ct) (Fig. 2B) . Similarly, no SARS-CoV-2 sequence was apparent on Sanger chromatograms for the NTC condition. No spike-in sequence was qualitatively discernable at the 5000 GCE dilution. Mixed bases were obviously present for both the 10 GCE and 100 GCE conditions suggesting that the limit of detection is about 10 GCE SARS-CoV-2.
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