ABI Prism® 7900HT Sequence Detection System ... - OpenWetWare

Programming a
Temperature Ramp
for Dissociation
Curve Analysis
4-16 Run Setup and Basic Operation
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 page 6-18 for a detailed explanation).
To add a temperature ramp to the method for dissociation curve analysis:
Step Action
1 From the Instrument tab of the plate document, click the Thermal Profile tab.
The software displays the Thermal Profile tabbed page.
2 Click Add Dissociation Stage.
The SDS software inserts a temperature ramp at the end of the thermal profile
consisting of a set of default steps.
The default temperature ramp can be customized, however Applied Biosystems
recommends the following guidelines to ensure the greatest separation of the
derivative peaks during the analysis, and therefore the maximum resolution for the
run.
Guideline Example
TheStartandEndstepsofthe
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 %.
Adatacollectioniconmustbe
placed on the temperature ramp in
the thermal profile.
3 Go to “Step 5 – Saving the Plate Document as a Template” on page 4-17.
End step
Start step
End step
ramp rate
setting
Data
collection
icon