The difference between the maximum and minimum annealing temperatures is commonly
between 5°C and 10°C starting with a maximum temperature that is 2°C above the higher
primer melting temperature (T
m
). For example, if the T
m
of Primer 1 is 60°C and the T
m
of
Primer 2 is 54°C, set the max temperature to 62°C and the min temperature to around 52°C.
4PCR Programs
Storage/Pause Step
50 Agilent SureCycler 8800 Se
tup and User’s Guide
Hold For Enter the duration of the annealing step.
Elongation
T
emperature and
Hold Ti
me
Enter the elongation temperature (in the Elongation Temperature field)
and the duration of elongation (in the adjacent Hold For field) to be used
during touchdown cycling.
Cycles Enter the number of cycles for the To
uch
down step.
Storage/Pause Step
The Storage/Pause program step cools the thermal block down to a
temperature between 4° and 12°C for a specified period of time or
indefinitely.
Hold Temperature Enter the storage temperature in °C.
Hold for Select this radio button if you prefer to maintain the temperature for a set
duration
of time. Ente
r the duration in the accompanying field.
Infinite Select this radio button to hold the specified store temperature for an
un
limit
ed period of time.
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