Adjusting The Baseline And Threshold - Applied Biosystems 7500 Getting Started Manual

Adjusting the Baseline and Threshold

The SDS Software 1.3.1 calculates baseline and threshold values for a detector based on
Automatic
the assumption that the data exhibit the "typical" amplification curve.
Baseline and
Threshold
A typical amplification curve has a:
Determination
Experimental error (such as contamination, pipetting errors, and so on) can produce data
that deviate significantly from data for typical amplification curves. Such atypical data
cause the software algorithm to generate incorrect baseline and threshold values for the
associated detector. Therefore, Applied Biosystems recommends reviewing all baseline
and threshold values after analysis of the study data. If necessary, adjust the values
manually as described on
If you set the baseline and threshold values manually for any detector in the study, you
Manual Baseline
must perform the adjustment procedure on
and Threshold
Determination
The following amplification plots show the effects of baseline and threshold settings.
Notes
Applied Biosystems 7300/7500/7500 Fast Real-Time PCR System Relative Quantification Getting Started Guide
• Plateau phase (a)
• Linear phase (b)
• Exponential (geometric phase) (c)
• Background (d)
• Baseline (e)
Threshold
∆R
n
Cycle
page
Chapter 6 Analyzing Data in an RQ Study
Adjusting the Baseline and Threshold
68.
page 68 for each of the detectors.
a
b
c
d
e
6
65