Applied Biosystems 7900HT User Manual page 185

Overview
Algorithmic
The SDS software can analyze raw data immediately after an absolute quantification
run is complete. The term raw data refers to the spectral data between 500 nm to
Manipulation of
660 nm collected by the software during the real-time run. During the analysis, the
Raw Data
software automatically applies several mathematical transformations to the raw data
to generate a more direct measure of the relationship between the spectral changes in
the unknown samples.
Multicomponenting
The first mathematical transformation involves the conversion of the raw data,
expressed in terms of Fluorescent Signal vs. Wavelength, to pure dye components
using the extracted pure dye standards. At the same time, the software determines the
contribution of each dye in the raw data using the multicomponent algorithm.
Setting the Threshold and Calling C s
T
After normalization, the baseline and threshold values must be set for the run
(see "Kinetic Analysis/ Quantitative PCR" on page D-4 for more information). The
results of the experiment can be visualized in the Standard Curve graph of the
Results tab. The graph consists of a scatterplot of standard and unknown samples
graphed on a linear-scale plot of Threshold Cycle (C ) versus quantity value.
T
DRAFT
Applied Biosystems 7900HT Fast Real-Time PCR System and SDS Enterprise Database User Guide 6-7
September 1, 2004 11:39 am, CH_Real-Time.fm