Real-time PCR reactions are the default setting for the Open qPCR system, but the instrument is also capable of functioning as a conventional/endpoint thermocycler by making a few adjustments in the protocol editor while setting up an experiment.
How to use Open qPCR for conventional PCR:
After creating a new experiment, the default protocol appears and the default real-time setting is indicated by the amplification icon (green arrow):
To switch to endpoint reactions, turn off Gather Data (GATHER DATA) for all steps & ramps in all stages of the protocol.
Toggle the STEP & RAMP options from the green “ON” position:
to the gray “OFF” position:
Confirm that the setting has been changed from real-time to endpoint by verifying that there are no amplification icons visible for any of the steps in the protocol.
Consequences of switching to conventional PCR
Choosing to turn off the data gathering functions will cause a few other changes that are important to keep in mind when deciding if switching to endpoint reactions is the right option. Using real-time settings, the results screen displays a generated amplification curve and Cq values if a dye or probe is used in the reaction.
Using conventional PCR reactions with no fluorescence detection, the LEDs will not be activated throughout the duration of the protocol. As a result, no amplification curve or Cq values will be generated (if using a dye or probe) and these areas will be blank on the results screen.