Software, Version 2.1 Owner's manual

Section 4 Workshop Module
- 37 -
Fig. 4.16. Melt Curve Protocol Editing Table.
6. Anneal curves can be also created by entering the temperature change as a negative
value (for example, –0.5).
7. Save the protocol by entering a file name and then clicking Save & Exit Protocol Editing.
NOTE: The minimum dwell times necessary for data collection are 10 seconds for 1
fluorophore. We recommend using a slightly higher dwell time than the minimum values
so that more data points are collected at each repeat.
4.3.6 Sample Protocol Files
There are several sample protocol files that can be used as editable templates for custom
protocol design, including:
• 2Step.tmo: This protocol can be used for most single fluorophore or SYBR
®
Green I
real-time PCR experiments
• 2StepAmp+Melt.tmo: This protocol can be used for most SYBR
®
Green I real-time
PCR experiments. The melt curve that follows the qPCR experiment can be used to
determine the number of amplicons produced in the qPCR reaction
• Gradient.tmo: This protocol can be used to determine the most optimal temperature to
perform real-time PCR experiments
4.4 Run Set
A Run Set is a linked pairing of a Plate Setup and Protocol files. Run Set files generated by the
iQ5 software are stored with the extension .run. Run sets are useful for experiments that are
repeated on a frequent basis.
When the Run Set tab is selected, run set files are shown in the file browser.
The protocol defined in the run set is shown in the Selected Protocol pane and the plate setup
defined in the run set is shown in the Selected Plate Setup pane (Figure 4.17).