Operator`s manual

35
Performing Gene Scanning Analysis
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Normalization ►
Click the Normalization tab to normalize the melting curves.
Fluorescence values acquired during High Resolution Melting will vary in magni-
tude, e.g., due to differences in the starting amount of template in each sample.
This variability can mask differences between genotypes. Therefore the first step
in the analysis process is to normalize the data. In normalization, two regions of
each curve are selected (one before and one after the major transition); these
are arbitrarily defined, respectively, as 100% fluorescence and 0% (baseline)
fluorescence.
The upper graph, the Melting Curves graph, contains two pairs of movable vertical
sliders that correspond to Pre-melt Low and High temperatures (colored green) and
to Post-melt Low and High temperatures (colored blue). The grey area between the
sliders indicates the area used for normalization.
Below the Melting Curves chart you will see two groups of values that you can set:
The left group is used to define the Pre-melt Temperature Range, while the right group
is used to set the Post-melt Temperature Range.
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