Figure 11

The melting curve analysis of the PCR products at the end of the amplification was done by measuring the Eva Green fluorescence by slow heating at 0.5°Cs ^-1 increments from 70 to 95°C, with continuous fluorescence collection. Accordingly, a melting curve was generated at the end of the PCR amplification for monitoring the specificity of PCR reaction. It was found that a single peak (single product) at the expected melting temperature of PCR product, Tm 76-87°C, was observed while no significant premature peaks were found indicating that primer dimers artifacts or incorrect amplification products were minimal and providing further evidence on the specific detection of the target mRNA genes.