Applied Biosystems 7900HT Fast Real-Time PCR System and SDS ...

OverviewAlgorithmicManipulation ofRaw DataThe SDS software can analyze raw data immediately after an absolute quantificationrun is complete. The term raw data refers to the spectral data between 500 nm to660 nm collected by the software during the real-time run. During the analysis, thesoftware automatically applies several mathematical transformations to the raw datato generate a more direct measure of the relationship between the spectral changes inthe unknown samples.MulticomponentingThe first mathematical transformation involves the conversion of the raw data,expressed in terms of Fluorescent Signal vs. Wavelength, to pure dye componentsusing the extracted pure dye standards. At the same time, the software determines thecontribution of each dye in the raw data using the multicomponent algorithm.Setting the Threshold and Calling C T sAfter normalization, the baseline and threshold values must be set for the run(see “Kinetic Analysis/ Quantitative PCR” on page D-4 for more information). Theresults of the experiment can be visualized in the Standard Curve graph of theResults tab. The graph consists of a scatterplot of standard and unknown samplesgraphed on a linear-scale plot of Threshold Cycle (C T ) versus quantity value.Applied Biosystems 7900HT Fast Real-Time PCR System and SDS Enterprise Database User Guide 6-7DRAFTSeptember 1, 2004 11:39 am, CH_Real-Time.fm