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Supplementary Figure S7: Improved SARS-COV-2 PCR detection and genotyping with double-bubble primers

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posted on 14.09.2021, 09:10 by Menachem Ailenberg, Andras Kapus, Ori D Rotstein

Using D-B primers to detect SARS-COV-2 virus in nasopharyngeal samples using fast conditions. (A) cDNAs from reverse-transcribed RNA extracted from nasopharyngeal swabs of 3 patients (S1, S2, S3) as well as SARS-COV-2 synthetic RNA “N” were subjected to qPCR using primer mix #8 and VIC-TqM probe #12 and TqM fast kit+ UDG. Ct values: Patient S1- 20.44, patient S2- 21.26, patient S3- 21.38, synthetic RNA- 25.78, NYC- 37.63. (B) one-tube RT-qPCR using template RNA extracted from nasopharyngeal swabs of positive patient S2 as well as SARS-COV-2 synthetic RNA “N”, TqM fast kit without UDG and iScript reverse-transcriptase. Insert- 1.5% agarose gel: lane 1- 100 bp ladder; lane 2- SARS-COV-2 synthetic RNA “N”. lane 3- RNA extracted from nasopharyngeal swabs of patient S2. Ct- values: Patient S1- 23.47, synthetic RNA- 27.36, NTC- undetermined.

Funding

This study was supported by a grant from the St. Michael’s Hospital Foundation (OR, AK) and a grant from CIHR # 400642 (OR).

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