High Resolution Melt (HRM) App - FAQs

What is the difference between continuous melt curves and step and hold melt curves on the Applied Biosystems Real-Time PCR Instruments?

Continuous melt curve: increases the temperature by the ramp increment (degrees C/sec) so that the temperature will change at a constant rate set in the protocol.
Step and Hold melt curve: increases the temperature by the ramp increment (degrees C) and then holds at that temperature for the time specified by the user.

The amount of data collected will depend on the filter(s) selected. The more filters you use, the longer the acquisition will take and the fewer data points will be collected.

Find additional tips, troubleshooting help, and resources within our Real-Time PCR and Digital PCR Instruments Support Center.

How can I create a new variant call group in the HRM app on Thermo Fisher Connect, select a color for the variant, and assign wells to that group?

To create a new variant call group in the HRM app on Thermo Fisher Connect, follow these steps:

1. Select the 'Quality Control & Results' tab.
2. In the 'Quality Control & Results' pane, select your experiment of interest.
3. Select the 'Derivative melt plot'.
4. Click 'Actions', and then select 'Manual Call'.
5. From the 'Manual Call' dialog box, you have two options:
1. To assign the sample to an existing variant call: click 'Select Existing', choose the appropriate call from the 'Group' drop-down menu, and then click 'OK'.
2. To create a new variant call: click 'Create New', enter a name for the new call in the 'Group' field, select a color, and then click 'OK'.
6. Repeat the above steps to assign more manual calls.
7. Click 'Analyze' to reanalyze the data using the manual calls.