BIO RAD CFX384, Instruction Manual

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CFX96 and CFX384 Systems Manual
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Table 32 lists the type of information shown in the RFU spreadsheet.
-d(RFU)/dT Spreadsheet
Select the -d(RFU)/dT spreadsheet to view the type of data shown in Figure 92.
Figure 92. The -d(RFU)/dT spreadsheet in the Melt Curve Data tab.
Table 33 lists the type of information shown in the -d(RFU)/dT spreadsheet.
End Point Tab
Open the End Point tab to analyze final relative fluorescence units (RFUs) for the sample wells.
The software compares the RFU levels for wells with unknown samples to the RFU levels for
wells with negative controls, and “calls” the unknown as a Positive or Negative. Positive
samples have an RFU value that is greater than the average RFU value of the negative controls
plus the Cut Off Value.
To analyze the end point data, the plate must contain negative controls, or the software cannot
make the call. Run one of these two types of protocols:
• Run a Quantitation protocol. Set up a standard protocol. After running the experiment,
open the Data Analysis window, adjust the data analysis settings in the Quantitation tab,
and then click the End Point tab to pick an end point cycle
• Run an End Point Only protocol. Load the End Point Only protocol in the Plate tab of
the Experiment Setup window, select or create a plate, and run the experiment
Table 32. RFU spreadsheet content
Information Description
Well number (A1, A2, A3, A4,
A5...) Well position in the plate for the loaded wells
Temperature Melting temperature of the amplified target. Plotted as one
well per row, and multiple wells for multiple products in the
same well
Table 33. -d(RFU)/dT spreadsheet content
Information Description
Well number (A1, A2, A3, A4,
A5...) Well position in the plate for the loaded wells
-d(RFU)/dT Negative rate of change in RFU as temperature (T) changes