It is indicated that RAPD reactions are made with a layer of mineral oil on the top of the mixture, iI would like to ask which oil to use and how and why it is used?
I actualy did the experiment, and I found after the reaction that the mixture was in the top of the tube (in the gel I had no bands)! is that because of the temperatures used during the PCR and is that the reason of using mineral oil, any detailed protocol will help!
Dear Amal,
If you use a PCR machine with a heated lid (99% of the contemporary PCR machines have a lid) then you can exclude the mineral oil. Just make sure your heated lid is on during the whole reaction (the lid should be on a temperature at 99-105C. Check the option "pre-heat lid").
Best regards,
Yehu
HI, the new generation of cylers all have a heated lid. How old is your cycler and which company etc.? Cheers, Nadine
Hi
You use mineral oil when your PCR machine has not heated lid. The benefit of oil is to prevents loss of liquid and changes in salt and buffer composition due to evaporation and condensation, might affect enzyme activity.
Thank you everyOne, i think that our machine (from BioRad company) has a heated lid , but i need to check the program and make sure i put the step of heating the lid, this was very helpful thank you again
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