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

To set the baseline and threshold values for the run: (continued)
Step Action
5 From the Plot drop-down list, select ∆Rn vs. Cycle.
The software plots the analyzed data in the graph of normalized reporter
fluorescence (∆Rn ) versus cycle number.
6 Identify the components of the amplification curve and set the threshold so that it is:
♦ Above the background
♦ Below the plateaued and linear regions of the amplification curve
♦ Within in the geometric phase of the amplification curve
R n
Cycle
Plateau phase
Linear phase
Geometric phase
Threshold setting
(click and drag)
Background
Baseline
7 Set the baseline and threshold for any remaining detectors present on the plate as
follows:
a. From the Detector drop-down list, and select another detector.
b. Repeat steps 3 to 6 until the baseline and threshold values have been set for
each detector.
8 Eliminate outliers from the analyzed run data as explained on page 6-12.
Real-Time Analysis 6-11