Software User Manual

Precision Melt Analysis Software Instruction Manual
33
5 Analyzing Melt Data
Read this chapter for information about analyzing melt data in Precision Melt Analysis
software.
Processing melt data (below)
Precision Melt tab (page 33)
Precision Melt Data tab (page 42)
Melt Curve tab (page 46)
Melt Curve Data tab (page 48)
Processing Melt Data
Precision Melt Analysis software plots the relative fluorescence unit (RFU) data collected
during a melt curve as a function to temperature. The software automatically starts with the
raw melt curve data and proceeds with the following steps:
Negatives Detection. All wells with sample content type designated NTC or Negative
Control in the Plate Editor are automatically considered negatives. Any well with a low
starting RFU is also considered a negative. All wells designated as negative are
automatically excluded from cluster analysis
NOTE: Override a well's automatically determined negative status by manually
including or excluding wells from cluster analysis. This can be done by right-
clicking on the well data in one of the charts, or through the drop-down selector for
the well in the data spreadsheet
TIP: Multiple wells can be called at once by holding down the right mouse button
and dragging over the wells to select multiple wells
RFU Normalization. All non-negative wells are normalized along the RFU axis (y-axis)
such that the average data value at the start of the pre-melt range is one, and the
average data value at the end of the post-melt range is zero
Clustering. Precision Melt Analysis software automatically determines a cluster
assignment for each non-negative well
Generate Difference Curves. For easy visual identification of clusters, the software
generates a Difference Curve chart of the data. The Difference Curve shows the
difference in fluorescence between a well and the fluorescence of a reference curve. The
reference curve is derived from the average fluorescence of all the curves within a
selected reference cluster