Open qPCR for isothermal amplification

#1

I would like suggestions on the best way to setup the Open qPCR device for only one 30 minutes cycle at a constant temperature.

I managed to get the amplification as expected by setting one cycling stage with only one cycle at the desired temperature. However this approach is kind of end-point while I would like to get also the amplification curve.

Since the amplification curve is based on the number of cycles and not time, would it be feasible to define a certain number of stages, each of 1 cycle and then obtain the amplification curve as an aggregate? Has anyone used the device for a similar application?

Thanks in advance.

Marco

#2

Hello @marco,

You’re definitely able to obtain a fluorescent read-out on your isothermal amplification protocol. Rather than setting one cycle for 30 minutes, you could set 60 cycles for 30 second increments each (30 minutes total). Make sure you have the “gather data” turned on for “step”. I’ve attached a screenshot of this example protocol at constant temp for your reference.

Cheers,Isothermal%20Protocol_Example
Lily

#3

Thank you very much. It worked perfectly!