Configuring a Temperature Step

A temperature step ramps to and holds a target temperature for a specified duration. You can configure a temperature step for the following block types:

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96-well

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96-deepwell

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384-well

Note: Temperature steps can include a plate read (fluorescence acquisition) at the end of the step.

Temperature steps appear in the graphical display as shown below.

To add or edit a temperature step

1.

Verify or change the existing values. The fields are explained in the following table.

Parameter	Description	Range
Temperature	Target temperature for the step	0.0°–100.0° C, in tenths of a degree
Time	Hold time for the step, in seconds; the hold time is the duration after the block has reached the target temperature	1–64800 sec
Increment	(Optional) the degrees to increase or decrease the step's temperature with each successive cycle Note: The increment is automatically set to 0 if the field is empty.	-10 to 10° C per cycle, in tenths of a degree
Ramp Rate	(Optional) instructs the thermal cycler to ramp to the target temperature at the specified rate Note: The ramp rate is automatically set to the maximum if the field is empty.	● 96-well: 0.1–5° C per sec ● 96-deepwell: 0.1–2.5° C per sec ● 384-well: 0.1–2.5° C per sec
Extend	(Optional) the amount of time to increase or decrease the step's hold time with each successive cycle. Note: The Extend time is automatically set to 0 if you leave the field empty.	-60 to 60 sec per cycle
Beep	When set to On, a beep sounds from the instrument when it reaches the target temperature.	On or Off
Read	When set to On, the CFX instrument executes a plate read (fluorescence acquisition) at the end of the step.	On or Off

2.

Click Add Step or Save Changes.

The temperature step is added or changed

Note: The Increment, Ramp Rate, Extend, and Beep parameters appear in the protocol step details, but not in the graphical display.

3.

Do one of the following:

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To add another step to your protocol, and the step and select the step type. For information, see the Help page for the specified step type.

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If you are finished creating or editing the protocol, click the Add button or Save button, respectively. The Manage CFX Protocols page opens, displaying the protocol and the creation or modification date.