Figure 2.

Performance of Georgia Tech multiplex primers and probes in several commercially available master mixes. Shown is GT multiplex primer/probe performance in commercial TaqPath, TaqPath Multiplex, and TaqMan Fast Virus 1-Step master mixes. Commercial master mix identity had no detectable impact on performance of the GT-made multiplex primer/probe mix. Due to the proximity of FAM and HEX channels, bleed-through from the FAM into the HEX channel was observed (see bottom nCov plasmid panels) but was of lower intensity than signal generated by the HEX-RP-BHQ1 probe (see top panels) and did not interfere with analyses when the HEX fluorescence threshold (blue dashed line) was set above the bleed-through intensity. Template in the top row consisted of synthetic SARS-CoV-2 RNA (ATCC) mixed with HEK293T RNA. Results are consistent with those expected for a positive patient sample. A negative sample would consist of a single amplification curve in the HEX channel (blue line). Template in the bottom row was 2019_nCoV_N_Positive Control (IDT) plasmid DNA.