Figure 2.

Real-time PCR for F5 Leiden, DNA extraction free method versus purified DNA method.

Notes: (A) Concentration range of WBCs (60×10⁶–0.2×10⁶ WBC/mL) show that the most suitable concentration of WBC for PCR and melting analysis purposes was located between 0.2–5×10⁶ WBC/mL. (B) Real-time CP values for the WBC versus purified DNA: CP mean value for WBC was 29.26±0.566 versus purified DNA 24.79±0.56. PCR from WBC versus purified DNA show a delay of about four cycles in CP.

Abbreviations: PCR, polymerase chain reaction; WBCs, white blood cells; CP, crossing point; PBS, phosphate buffered saline; −(d/dT), negative derivative of fluorescence over temperature.