Operator`s manual

31
Performing Gene Scanning Analysis
Performing Gene Scanning D
Analysis
You can perform a Gene Scanning analysis on any experiment that contains a melting
curve program. The LightCycler® 480 Gene Scanning Software determines the heterodu-
plex structures in samples by analyzing experimental data generated in the presence of
the LightCycler® 480 High Resolution Melting Dye. After samples are amplified by PCR
and subjected to a melting curve experiment, the software analyzes the shapes of the indi-
vidual curves and groups samples that have similar melting curves.
You can also include melting standards of known sequence variants or genotypes in your
experiment. In this case, the software compares the melting curves of the individual sam-
ples to the designated in-run melting standards.
Without the LightCycler® 480 Gene Scanning Software you can still view any gene
scanning experiment in the LightCycler® 480 Software but you cannot perform a Gene
Scanning analysis.
To perform a gene scanning experiment:
�
Set up an experiment containing an amplification program (to amplify the target
samples) and a melting curve program (to melt the samples). Whether you need to
include an internal standard in your experiment depends on the analysis mode you
want to use:
Auto Grouping or Common/Variants analysis: no melting standards are required ►
In-run melting standards: include melting standard samples ►
If you wish, you may also include control samples (e.g., negative controls or no-tem-
plate controls) in your experiment.
Internal standards act as positive controls.
► ► ►