Viral RNA was extracted from tissue samples using QIAmp Viral extraction kit following the manufacturer’s instructions. RT-qPCR was performed according to an established protocol34 (link),37 (link). Primers and probe targeting SARS-CoV-2 E gene were used following the Super Script III Platinum One-Step RT-qPCR (Invitrogen) protocol. Amplification was performed as follows: reverse transcription 55 °C 20 min, denaturation 95 °C 3 min, followed by 50× cycles of amplification at 95 °C 15 s, 58 °C 30 s, where data was acquired. Further analysis and Cq values were determined using the BioRad CFX Maestro software (BioRad).
Kaiser F.K., Hernandez M.G., Krüger N., Englund E., Du W., Mykytyn A.Z., Raadsen M.P., Lamers M.M., Rodrigues Ianiski F., Shamorkina T.M., Snijder J., Armando F., Beythien G., Ciurkiewicz M., Schreiner T., Gruber-Dujardin E., Bleyer M., Batura O., Erffmeier L., Hinkel R., Rocha C., Mirolo M., Drabek D., Bosch B.J., Emalfarb M., Valbuena N., Tchelet R., Baumgärtner W., Saloheimo M., Pöhlmann S., Grosveld F., Haagmans B.L, & Osterhaus A.D. (2024). Filamentous fungus-produced human monoclonal antibody provides protection against SARS-CoV-2 in hamster and non-human primate models. Nature Communications, 15, 2319.
Other organizations :
University of Veterinary Medicine Hannover, Foundation, German Primate Center, VTT Technical Research Centre of Finland, Utrecht University, Erasmus MC, Harbour BioMed (Netherlands)
RT-qPCR protocol using Super Script III Platinum One-Step RT-qPCR (Invitrogen)
Primers and probe targeting SARS-CoV-2 E gene
dependent variables
Viral RNA levels (as indicated by Cq values)
control variables
Manufacturer's instructions for QIAmp Viral extraction kit
Established RT-qPCR protocol34,37
Amplification conditions: reverse transcription 55 °C 20 min, denaturation 95 °C 3 min, followed by 50× cycles of amplification at 95 °C 15 s, 58 °C 30 s
Analysis using BioRad CFX Maestro software
positive controls
Not explicitly mentioned
negative controls
Not explicitly mentioned
Annotations
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