Touchdown - Agilent Technologies SureCycler 8800 Setup And User Manual

Touchdown

Touchdown PCR is an advanced PCR technique used to reduce nonspecific
primer/template binding. In the early cycles of touchdown PCR, the
annealing temperature is set relatively high to promote specific
amplification. Then, in later cycles, the annealing temperature is lowered
to permit more robust amplification.
The Touchdown program step is provided as a one- step command for easy
programming of touchdown PCR. You enter the maximum (starting)
annealing temperature and the minimum (ending) annealing temperature.
Then, during the run, the software executes the cycling protocol so that
the annealing temperature is lowered by an equal amount with each cycle.
Denaturation Enter the denaturation temperature (in the Denaturation Temperature
Temperature and field) and duration of denaturation (in the adjacent Hold For field) to be
Hold Time used during touchdown cycling.
Max Annealing Enter the maximum annealing temperature. The first cycle of the PCR
Temperature program will use this annealing temperature.
Min Annealing Enter the minimum annealing temperature. The last cycle of the PCR
Temperature program will use this annealing temperature.
Agilent SureCycler 8800 Setup and User's Guide
PCR Programs 4
Touchdown
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