Does a touchdown PCR protocol improve results?
No. Relative peak areas of the probe amplification products change depending on the PCR protocol used. Even though each fragment is amplified with the same PCR primers, some fragments have stronger reduced amplification efficiencies at annealing temperatures in excess of 64°C. The annealing temperature and the speed of temperature changes are often not perfectly uniformly distributed across the different parts of the thermocycler. The reproducibility of results may therefore be lower when the first PCR cycles have a low efficiency.
Related Questions
- In March 2006, three labs published their results using the Faustman labs protocol for end-stage diabetes reversal in mice. What were the results?
- Why is the QuantiFast denaturation step different for PCR and RT-PCR runs in the two-step protocol for the ABI 7500 and other cyclers?
- Does a touchdown PCR protocol improve results?