PCR Programs 4
To create a PCR program using the PCR Wizard
Agilent SureCycler 8800 Setup and User’s Guide 41
5 Ad
justments
• Select adjustments t
o
the program.
•“GC” content larger th
an 70%? If the DNA template contains
greater than 70% GC base pairs, add this adjustment. The wizard
will adjust the PCR program to compensate for the higher melting
temperature of the template.
•Do you want a gradient annealing step? If y
ou want the annealing
step of the PCR program to be a temperature gradient, add this
adjustment. The wizard will determine an appropriate gradient
range based on the melting temperatures of the primers.
Once the wizard has guided you through these five steps, it creates a PCR
pr
ogram
based on the provided information and displays it in the
programming screen with the steps of the program displayed in order in
the Program Window.
At this point, you may add, edit or delete steps in the same way as you
would for a manually created program (see “Editing PCR Programs” on
page 53).
To view or edit the parameters of any step, double- press the step in the
Program Window. A new screen will open showing the settings for that
command, such as temperature and duration. See the section “Program
Steps in the Commands List” on page 43 for more information on the
parameters for each command.
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