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

To create a method for the absolute quantification run: (continued)
Step Action
3 Modify the default thermal profile for the method as needed:
To… Then…
adjust step
parameters
select a text field value, type a new value, and press Enter.
(time/temp)
Temperature text field (between 4 and 99.9 °C)
add a hold,
cycle set, or
step
Time text field (between 0:01 and 98:59 minutes)
a. Click step to the left of the location you want to place the
new stage.
b. Click Add Cycle or Add Hold.
The software inserts the stage into the thermal profile.
Note Toaddasteptoastage,selectthesteptotheleftof
the location you want to place the step and click Add Step.
remove a step a. Click the step you want to remove.
The software highlights the selected step.
b. Click Delete Step to remove the step from the profile.
4 Configure the data collection options for the method as follows:
a. Click the Data Collection tab.
b. Click each plateau or ramp within the cycle stage of the thermal profile to place a
data collection icon at each step.
Data collection icons
Selected
step
New step
appears here
5 Click the Sample Volume (µL) text field and type the volume of the reactions to be
run on the plate.
Note ‘Sample Volume’ refers to the entire contents of any well, including buffer
blank, or any combination of master mix and nucleic acids.
IMPORTANT All wells on one plate must contain the same reaction volume.
6 Choose from the following options:
If performing an… Then…
absolute quantification run
only
absolute quantification run
with a dissociation curve
go to “Step 5 – Saving the Plate Document as a
Template” on page 4-17.
add a temperature ramp to generate dissociation
curve data as explained on page 4-16.
Run Setup and Basic Operation 4-15