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-   -   Vapor in PCR after using thermocycler? (http://www.molecularstation.com/forum/pcr-polymerase-chain-reaction-forum/88214-vapor-pcr-after-using-thermocycler.html)

brasarehot 04-01-2014 12:21 AM

Vapor in PCR after using thermocycler?
 
After my PCR is run for X number of cycles I see these droplets on the sides of my PCR tube above my PCR solution. I was wondering what this is and is it normal/preventable? I am worried since there are droplets above the solution that the entire solution didn't go through PCR. I have it set up to preheat lid and it's on simulated tube option. I am using PCR Sprint thermocycler.

I am new to PCR so thank you for your help

butters 04-01-2014 03:45 AM

Re: Vapor in PCR after using thermocycler?
 
if you can put the picture is great.

I guess it not really biggy generally i do see them especially if I do not use mineral oil.

luisillo 04-15-2014 04:30 PM

Re: Vapor in PCR after using thermocycler?
 
Do you add an extra cooling cycle after PCR ends (i.e. 4C for 10 min)? I usually add this cycle since it gives enough time to the tube and lid to cool down, so you avoid vapors and burns, respectively. Droplet formation in this scenario has proven to be harmless to PCR product results in my case.

If you do not use this last step, condensation at the top of the tube might be caused by the sudden drop in temperature in the top of the tubes when you open the lid (>100 C) and expose them to room temp (~20 C). Similar to what happens when using the 4C cycle.

At any rate, droplets should be no big deal for your PCR.


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