Bio-Rad iQ™5 Optical System Software, Version 2.1 - Manual (Page 83)

Section 6 Data Analysis Module
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Fig. 6.34. The Select Cycle Drop-Down List Box.
6.7.7 Vertical Threshold
The Vertical Threshold box (Figure 6.35) displays the current position of the vertical bar. This box
is not editable. The positions of the vertical bar along with the position of the horizontal bar
determine the genotype assignment for unknown samples.
Fig. 6.35. The Vertical Threshold Box.
6.7.8 Horizontal Threshold
The Horizontal Threshold box (Figure 6.36) displays the current position of the horizontal bar.
This box is not editable. The position of the horizontal bar along with the position of the vertical
bar determines the genotype assignment of unknown samples.
Fig. 6.36. The Horizontal Threshold Box.
6.7.9 Normalize Data
When the RFU option button is selected, the Normalize Data option (Figure 6.37) appears as
shown below if the plate setup contains a no template control (NTC) sample type for both Allele
1 and Allele 2.
Fig. 6.37. The Normalize Data Button.
The RFU data may be normalized and displayed on a plot that ranges from 0 to 1 on both axes.
This display is sometimes a very effective presentation of RFU data. The RFU data is normalized
to the NTC value as a linear combination of Allele 1 and Allele 2 specific signals. The iQ5 software
uses the following formula initially introduced by Livak et al. (1995):