Bio-Rad CFX Manager Quick Guide

AMPLIFICATION

CFX Manager™ Software
Protocol Quick Guide

Experiment Setup Protocol Tab

The Protocol tab displays a preview of the protocol
loaded in the Experiment Setup window (Figure 1).

Click Create New to open the Protocol Editor
to create a new protocol.

Click Select Existing to launch the file browser
to load a protocol to use in an experiment or to edit.

Use the Express Load drop-down menu to directly
load a protocol to use in an experiment or to edit.

Click Edit Selected to open the Protocol Editor
to edit the steps of the selected protocol.

Click the Start Run tab to proceed and run an
experiment with the currently loaded protocol.

Protocol Editor

The Protocol Editor is used to create a new protocol
or edit an existing one (Figure 2).

1. Select any step in either the graphical or text
display — the step becomes highlighted in blue.
Click on the temperature or dwell time to directly
edit the value.

2. Click Insert Step to add a temperature step to
the protocol.

Click Delete Step to remove a highlighted

step from the protocol.

3. Click Add Plate Read to Step to designate
when fluorescence data will be acquired during
the protocol. The text on this button changes to
Remove Plate Read if the currently highlighted
step has a plate read. If you do not want to
acquire data at that step, then click Remove
Plate Read.

4. Click on the number of repeats of a GOTO Step
to change the number of cycles in the protocol
(Figure 2, Step 4).

Fig. 1. Protocol tab in the Experiment Setup window. Load an existing protocol or
create a new protocol for an experiment.

Graphical
display

Text display

Protocol
editing
controls

Fig. 2. Protocol Editor. The protocol editing controls are on the
left. Both the graphical and text displays can be edited. Step 1 is
highlighted and 95˚C is selected for editing in the text display.

Click on the GOTO Step number to change

the steps included in the GOTO loop.

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Group

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To add a gradient step:

1. Click Insert Gradient in the protocol editing
controls (Figure 2).

2. To edit the temperature values for the lowest and
highest temperatures in the gradient, click on the
values in the graphical display, in the text display,
or in the gradient range calculator that appears
to the right of the text display (Figure 3).

3. Within the gradient range calculator, a specific
temperature can be assigned to any row.
Each row is then populated with the appropriate
temperature to satisfy the specialized range.

To add a melt curve:

1. Click Insert Melt Curve in the protocol editing
controls (Figure 2).

Graphical display

Fig. 3. Insert Gradient step in the Protocol Editor. A gradient step is
highlighted in blue in the graphical and text displays of the protocol. The gradient
range calculator appears when a gradient is added to the protocol.

Gradient range
calculator

2. In the graphical or text display, click on the
temperature values to edit the lowest and
highest temperatures of the melt curve range
(Figure 4, Step 5).

3. Click on the increment value to edit the
temperature interval at which data are acquired.

4. Click on the dwell time to edit the incubation
time for each temperature increment.

Fig. 4. Insert Melt Curve step in the Protocol Editor. A melt curve step is
highlighted in blue in the graphical and text displays of the protocol.

Bio-Rad’s real-time thermal cyclers are licensed real-time thermal cyclers under
Applera’s United States Patent No. 6,814,934 B1 for use in research and for all
other fields except the fields of human diagnostics and veterinary diagnostics.