Hello everyone

recently I got a qTOWER3 (a termocycler for qPCR) so, I do not have a lot of experience in this machine and I would ask you about set up a Melt curve. Firstly, I have a assay for detection of mutations of SARS-CoV-2 , according the insert show the following parameters (adjunt QTOWER3 PDF table 1) but the software of qTOWER3 just ask the following parameters (adjunt QTOWER3 PDF table 2), I have been triying to put in the termocycler the following data:

start: 40 °C

end: 75 °C

increment: 1°C

equilibration: 6 seconds (a doubt)

heating rate: 1.5 °C/sec

Graphically, The results that I have got are:

- Intervals of temperature (X axis) is according to the insert (40-75 °C)

- The problem is the formation of many curves (see QTOWER.jpg), for example for detection of positition 417 and 484 in gen spike, the insert show me the formation of two curves ( see MELT CURVE QTOWER )

I would to know if someone knows more about this termocycler (qPCRtouch) and how to create a DNA melt curve.

Any suggestions I will be very grateful.