Hi, I've been curious about the performance of a PCR machine, so I measured the temperature of its block with its lid open as it was running in 95 C for a couple of minutes. However, the temperature measured by (multiple) thermometers was below 80 C in various points on the block.
Is this thermal difference due to the opened lid, and hence it is normal? Or the PCR machine needs further calibration?
@Dirk Schmidt I agree with your answer, when he open the lid of the thermocycler constant temperature cant be obtained from each holes.
Otherwise is it Hg liquid thermometer?
You will never get the exact/reliable temperature using those kind of thermometers because it cant sense exact temperature of the site what you want....
It's most likely reading low because a standard thermometer doesn't reach to the bottom of the well the way a PCR tube does. Plus the lid being open lets a lot of the heat escape.
You can do a simple empirical test to see if it's working. Set up a PCR reaction that you know works well in another machine. You don't need to include negative controls since you are looking for presence of amplification. Put a good "positive control" DNA in every tube, put a tube in every well in the block, and see if you get amplification from all of the wells.
Good luck!
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