Applied Biosystems 7900HT User Manual page 107

Programming a
Temperature
Ramp for
Dissociation
Curve Analysis
Applied Biosystems 7900HT Fast Real-Time PCR System and SDS Enterprise Database User Guide
To generate the data required to perform a dissociation curve analysis, the 7900HT
instrument must be programmed to run a 'temperature ramp' in which it slowly
elevates the temperature of the samples while collecting fluorescence measurements
once every 7-10 seconds (see
1. In the Instrument tab of the plate document, select the Thermal Profile tab.
2. Click the step to the left of the location you want to place the new stage.
3. In the Thermal Profile tab, click
The SDS software inserts a temperature ramp at the end of the thermal profile
consisting of a set of default steps.
4. You can customize the default temperature ramp, however Applied Biosystems
recommends that you observe the following guidelines to ensure the maximum
resolution for the run (the greatest separation of the derivative peaks during the
analysis).
Guideline
The Start and End steps of
the temperature ramp must:
• be separated by a
minimum temperature
difference of 35 °C.
• elapse 15 seconds (0:15)
each.
The ramp rate setting for the
End step of the temperature
ramp must be 2 %.
IMPORTANT!
You must
deselect the 9600 Emulation
check box to modify the
Ramp Rate Settings.
The temperature ramp in the
thermal profile must contain
a data collection icon.
5. Go to "Step 6 – Saving the Plate Document as a Template" on page
Step 5 – Programming the Plate Document Method
page 6-38 for a detailed explanation).
Example
DRAFT
September 1, 2004 11:39 am, CH_Run.fm
.
End step
Start step
End step
ramp rate
setting
Data
collection
icon
3-24.
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