Agilent Technologies SureCycler 8800, Setup And User Manual

Page: 49 / 66

PCR Programs 4
Touchdown
Agilent SureCycler 8800 Setup and User’s Guide 49
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
Temperature and
Hold Time
Enter the denaturation temperature (in the Denaturation Temperature
field) and duration of denaturation (in the adjacent Hold For field) to be
used during touchdown cycling.
Max Annealing
Temperature
Enter the maximum annealing temperature. The first cycle of the PCR
program will use this annealing temperature.
Min Annealing
Temperature
Enter the minimum annealing temperature. The last cycle of the PCR
program will use this annealing temperature.