I use 0.2mL tubes for my PCR reactions and after each run there are always a few tubes that have less volume than the other tubes. There doesn't seem to be any pattern. I don't believe it is pipetting error. Does this happen to anyone else?
Hi Lacey,
When you first take your samples out of the machine, you should try and spin them down to see if they even out as some vapor can end up in the lid. Though if you are using a thermocycler with a heated lid, evaporation should not be an issue.
If you are using a heated lid and the samples are still evaporating, your tubes may not be closed all the way. Ensure that your tubes are tightly closed prior to PCR.
As a last resort, you could try adding 30 uL mineral oil to the top of the reaction mix to prevent any kind of evaporation. Good luck!
Hi Lacey,
When you first take your samples out of the machine, you should try and spin them down to see if they even out as some vapor can end up in the lid. Though if you are using a thermocycler with a heated lid, evaporation should not be an issue.
If you are using a heated lid and the samples are still evaporating, your tubes may not be closed all the way. Ensure that your tubes are tightly closed prior to PCR.
As a last resort, you could try adding 30 uL mineral oil to the top of the reaction mix to prevent any kind of evaporation. Good luck!
I agreed with answer above.
I suggest check and replace your tubes, maybe some of them damaged by pressure and autoclave or etc.
For those who add oil to the top, when you take out the sample to run on the gel do you pipette off the oil or how do you handle getting sample out?
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