pone.0243333.s001.tif (8.93 MB)

Titration of the diluted nucleocapsid spike-in transcript.

figure

posted on 2021-04-14, 17:55 authored by Julien Fassy, Caroline Lacoux, Sylvie Leroy, Latifa Noussair, Sylvain Hubac, Aurélien Degoutte, Georges Vassaux, Vianney Leclercq, David Rouquié, Charles-Hugo Marquette, Martin Rottman, Patrick Touron, Antoinette Lemoine, Jean-Louis Herrmann, Pascal Barbry, Jean-Louis Nahon, Laure-Emmanuelle Zaragosi, Bernard Mari

A ten-fold serial dilution ranging from 1 to 10–6 was prepared from the stock solution of the in vitro-transcribed N gene and supplemented with 2 ng/μL of total RNA from HEK 293 Cells. Reverse Transcription was performed followed by 15 cycles of pre-Amplification and 30 cycles of qPCR. The RT-qPCR reaction was performed without (A) or with (B) a RNA purification step. Linear regression was performed by logarithmic plots of transcript copy number against Cq value. We observed a good correlation according Cq linear regression curves according to dilution for the three viral CDC primers/probe sets (N1, N2, N3). No Cq value has been detected for the E primers/probe. RNP, used as internal control, shows constant detection of Cq value, suggesting a good performance of the qPCR.

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