What ramp mode should I use for HRM experiments using the QuantStudio 3 and QuantStudio 5 Real-Time PCR Systems?
We recommend using Continuous mode (set to 0.025 degrees C/sec) for HRM experiments. You can also choose “Number of Data Points per Degree”, but if you do so, make sure to collect at least 10 points per degree so that there is enough data for analysis. When in Continuous mode, the instrument uses a fixed ramp rate for the melt step and captures as many data points as possible within the time permitted. When in Data Point Per Degree mode, the instrument adjusts the step and hold for the melt step to achieve the specified number of data points.
Find additional tips, troubleshooting help, and resources within our Real-Time PCR and Digital PCR Instruments Support Center.
What calibrate plate can I use for High Resolution Melt (HRM) analysis on the QuantStudio 3 and QuantStudio 5 Real-Time PCR Systems?
We recommend using our MeltDoctor HRM Calibration plates. Use the plate type that matches the block on your QuantStudio 3 or QuantStudio 5 Real-Time PCR System:
0.1 mL, 96-Well: Cat. No. 4425618
384-Well: Cat. No. 4425559
0.2 mL, 96-well: Cat. Nos. 4425562 and 4425557 (Note: We do not have a 0.2 mL calibration plate, but you can make one by combining the MeltDoctor HRM Calibration Standard with the MeltDoctor HRM Reagent Kit)
Find additional tips, troubleshooting help, and resources within our Real-Time PCR and Digital PCR Instruments Support Center.
Are the QuantStudio 3 and QuantStudio 5 Real-Time PCR Systems compatible with High Resolution Melt (HRM) experiments?
Yes, you can perform HRM experiments on the QuantStudio 3 and QuantStudio 5 Real-Time PCR Systems. The data can either be analyzed in the HRM v3.1 Software (Cat. No. A30150), or in the Thermo Fisher Cloud HRM Analysis Application. Please see here (https://tools.thermofisher.com/content/sfs/manuals/MAN0014394_HighResMeltExperiment_GSG.pdf) for the user manual (with Cloud workflow).
Note: QuantStudio 3/QuantStudio 5 Real-Time PCR System firmware version 1.1 or later is required to perform HRM experiments.
Find additional tips, troubleshooting help, and resources within our Real-Time PCR and Digital PCR Instruments Support Center.
Where can I find information regarding High Resolution Melt (HRM) software v3.1 and HRM software for use on the QuantStudio 3 and QuantStudio 5 Real-Time PCR Systems?
For more information about HRM software v3.1 and HRM software for use on the QuantStudio 3 and QuantStudio 5 Real-Time PCR Systems, please go here (http://tools.thermofisher.com/content/sfs/manuals/MAN0014394_HighResMeltExperiment_GSG.pdf).
Find additional tips, troubleshooting help, and resources within our Real-Time PCR and Digital PCR Instruments Support Center.
