Bio-Rad Precision Melt Analysis™ Software - Manual (Page 38)

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