Operator`s manual

36
LightCycler® 480 Gene Scanning Software
Performing Gene Scanning Analysis
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LightCycler
®
480 Gene Scanning Software automatically places the temperature slid-
ers in a suitable region for normalization and displays the normalized data in the lower
graph (Normalized Melting Curves). Examine the upper graph to make sure that the
Pre-melt Temperature Range lies in an area where the background fluorescence of all
the samples is dropping consistently but no temperature transitions have occurred,
and that the Post-melt Temperature Range is placed in an area where melting is com-
plete for all samples.
If necessary, adjust the temperature settings by either dragging the vertical sliders
on the upper graph or by changing the values in the Slider Settings fields. For most
purposes, the recommended temperature interval between each set of vertical sliders
is about one degree.
If the Pre-melt and Post-melt Temperature Ranges are set correctly, you should then
obtain a set of normalized melting curves that have a flat section before and after
the major transition; no curve should rise very far above the upper line or fall very far
below the lower line.
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