Bio-Rad Precision Melt Analysis™ Software - Manual (Page 51)

Analyzing Melt Data
46
Difference RFU Spreadsheet
Select the Difference RFU spreadsheet to view the difference fluorescence readings for each
well. Negative wells do not appear in this spreadsheet because they are excluded from
difference processing (Figure 37).
Figure 37. Precision Melt data tab showing the Difference RFU data.
The Difference RFU spreadsheet includes the type of information shown in Table 21.
Melt Curve Tab
In the Melt Curve tab, Precision Melt Analysis software plots the RFU data collected during a
melt curve as a function of temperature. To analyze melt peak data, a beginning and ending
temperature is assigned to each peak. The floor of the peak area is specified by the position of
the melt threshold bar. A valid peak must have a minimum height relative to the distance
between a threshold and the height of the highest peak.
Open the Melt Curve tab (Figure 38) to determine the melting temperature (T
m
) of amplified
PCR products. This tab shows the melt curve data in these four views:
• Melt Curve. View the real-time data for each fluorophore as RFUs per temperature for
each well
• Melt Peak. View the negative regression of the RFU data per temperature for each well
• Well Selector. Select wells to show or hide the data
• Peak spreadsheet. View a spreadsheet of the data collected in the selected well
NOTE: This spreadsheet only shows as many as two peaks for each trace. To see
more peaks, click the Melt Curve Data tab (page 48).
Table 21. Difference RFU spreadsheet contents
Information Description
Well number
(A2, A3, A4, A5, A6...) Well data, listed by position in the plate for all the loaded wells
Temperature Temperature data listed in ascending order for all the loaded
wells