Today is going to be dedicated to getting temperature readings from the PCR machines (both OpenPCR and the ThermoCycler). Both machines have their own temperature output, but in the past I’ve discovered that they don’t really reflect the temp inside the PCR reaction tube (especially with regards to the ThermoCycler). The last time I did this with the ThermoCycler, I got some interesting results which led to my current program settings.
Today I’m going to repeat that experiment and replicate it with OpenPCR. Here is my setup:
- You will need:
- a PCR machine
- reaction tubes
- a thermocouple – These are pretty easy to find
- a temperature reader – I’ve used digital multimeters in the past, but for this experiment I’m using the TC-48-20 OEM because it comes with software that takes frequent measurements and allows me to export the data.
- Take a PCR reaction tube and drill a small hole in it
- Put either water or mineral oil in the tube (in the amount that you normally use for PCR reactions), I’m doing 50ul for this experiment.
- Put the thermocouple in the tube through the drilled hole and place the tube in the block on the PCR machine.
- Connect the thermocouple to your temperature reader (in my case I need to connect the temp probe to the TC-48-20 and then connect that to my computer)
- Run the PCR program and collect data.
Check out the images below:
