I would like to know whether some specific biomolecules will also evaporate and form vapors along with solvent vaporization.
If so, what kind of biomolecules?
If temperature of lid is 10 degree higher (105-110 degree) than denaturation (94-97 degree) step, there will be no evaporation.
But, if your reaction mixture evaporate, there are chances of evaporation of molecule like dNTPs, Mg, primers.
I think DNA may not evaporate.
Maybe not.
The cover of current PCR machines makes more heat above the 105-celsius degree, which can prohibit the condensation of water on the top of the PCR tubes during PCR cycles.
And some researchers add an oil-like solution (mineral oil) to prevent from vaporization of the PCR solutions.
If temperature of lid is 10 degree higher (105-110 degree) than denaturation (94-97 degree) step, there will be no evaporation.
But, if your reaction mixture evaporate, there are chances of evaporation of molecule like dNTPs, Mg, primers.
I think DNA may not evaporate.
Dear Krishnadas Narayanan Nampoothiri I agree with colleagues regarding hot lid in PCR machine and oil. of course large molecules are not very mobile and fly weakly. If we consider the genomic DNA. In the case of pDNA or amplicons, viral particles - I am not sure. I think that short molecules are able to fly.
Hi,
the energy you have during the PCR (max 98*C) is WAY to low to volatilize salts, or nucleotides, or DNA (including primers). They would rather decompose than evaporate. Decomposition products (ethanol, CO2, acetate, ammonia, etc.) may evaporate.
Thank You everyone for the answers. I appreciate them.
Andrei S. Babenka :Your comment on "short molecules are able to fly": May I know what kind of molecules and could you please pinpoint a few examples.
Jitendra Kumar Shandilya : Your comment on " But, if your reaction mixture evaporate, there are chances of evaporation of molecule like dNTPs, Mg, primers": Could you please provide some literature.
Dear Krishnadas Narayanan Nampoothiri thank you for your question.
If you open PCR tube after amplification (without mineral oil) you may receive contamination. I have found my amplicons (swab - elution - PCR) around the tube in rack and on the table. So I think that amplicons can fly. It was a real time PCR product. It was an accident...
When I worked with pDNA as a matrix - the same picture.
Of course somebody can say - sample handling or pipetting mistakes, rough and dirty work, contamination of reagents...but reagents were clear and I have ckecked all other points.
It is difficult for me to work clear with pDNA and amplicons...as a result )
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