qRT-PCR using SYBR Green I dye based detection approach. Real time PCR to determine the sensitivity and specificity of SARS-CoV-2 detection primers of N1, N2, N3 and C1 (RNase P internal control) using SYBR Green I dye. Series of dilutions of each synthetic DNA fragment (1pg, 10pg, 100pg and 1000pg) were used in the background of (a) MCF-7 cell line cDNA (10ng), (b) tongue squamous cell carcinoma (TSCC) sample genomic DNA (10ng), and (c) RNA synthesized by IVT in the background of MCF7 cell line RNA (50ng), for performing real-time PCR. For each primer, Ct values (in triplicates) in different concentrations of synthetic DNA/RNA fragments are plotted in the graph. Specificity of the primers to amplify single amplicon is represented as single peak in melting curves generated by “melting curve genotype analysis” in Roche Light cycler 480 machine.