Figure 2

Thermal denaturation (fluorescence extinction) profiles of the E6-2 amplicons generated from pHV101 and SiHa DNA. RFU, relative fluorescence units. (A) Family of curves obtained with serially diluted pHV101 or 50 ng of SiHa DNA. Threshold (thick horizontal line): 50 RFU. Curve 1, 2.5 × 10⁶ pHV101 copies. Curve 2, 2.5 × 10⁵ pHV101 copies. Curve 3, 2.5 × 10⁴ pHV101 copies. Curve 4, 2.5 × 10³ pHV101 copies. Curve 5, 2.5 × 10²pHV101 copies. Curve 6, Blank (without DNA). Curve 7, normal human blood DNA. Curve 8, 50 ng of SiHa cell DNA with the pU1M/pU1R primer pair. Curve 9, 50 ng of SiHa cell DNA with the pU1M forward primer only. (B) Thermal fluorescence extinction profiles of nested qPCR products. Profiles of pHV101 and SiHa DNA mixtures with both primers (peaks at 85.5°C), mixtures without DNA or with normal human blood DNA with both primers (peaks at 78.5°C), and SiHa DNA with the pU1M primer only (nearly horizontal line close to the baseline).