Say I'm doing something that's sensitive to temperature in a thermocycler. I need to have the "hot lid" on to prevent sample evaporation, but that's now delivering heat to my samples via the sample tube. The lid of our thermocycler is fixed at 105 degC. I'm worried that my samples will be warmer than the block-determined temperature.
I'm not familiar with all the intricacies of thermocyclers. Will my sample temperature be any higher than the block temperature if I have the "hot lid" on? Is this on the order of a degree? 10 degrees? Anyone have thoughts?
Ted, be sure your samples will be exactly at the temperature of the block. Your PCR reaction mixture is completely surounded by the block and separated by the microtube wall which is thick only 1mm or less. So, it is more likely that even if the tube is heated by the lid and the heat travels longitudinally downwards, wide comunication surface between a tube and the block will quickly and effectively regulate the mix to the set temperature.
These guys are correct..it will be the same temperature as the block. The lid needs to be higher in temperature to stop condensation occurring in the lid of your PCR tube. That way the concentration of all your reagents in your PCR mix remains stable during cycling. I know this for a fact because we sometimes use a temperature probe in a PCR tube to monitor the temperature in the block, (some PCR machines come with probes for monitoring temp.)...and the temp never seems to be affected by the lid.
I have a doubt ..what will happen if the lid temp maintained at 4 degrees while running PCR
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