Master mix consist of dna polymerase taq and other reagents such as nucleotides can it be negatively affected by exposure to the uv yet can eliminate the contamination?
Hi Asmaa Nasr. I think that way is not best choice. Because after uv exposure, the master mix will be trash. One hour in room temperature is a risk for enzymes in the mix. On the other hand, if you do not know the contaminants, it is will be waste time. Anything can be caused for this situation. The best way, order new one. Have a nice work.
Hi
I agree with huseyin. The UV irradiation will affect your enzymes as well as dNTPs. In best case you will not get any amplification afterwards. Even worse would be if you see an amplification for strongly expressed targets which let you think all is good but in the end you might loose low expressed targets thus generating wrong results.
I would agree to replace the reagents after thorough cleaning and repeat the experiments
Good luck Sven
I would not do that. For the reasons that Huseyin and Sven stated. And there's no way to guarantee that the contaminating DNA won't be picked up in your PCR reaction. Best thing to do is start fresh with all new reagents.
But before are you sure that the contamination is inside the master mix?
It is difficult process since uv could not eradicate your contaminant selectively. It will deactivate all of functional units for PCR. If you are curious and have time try it!
Try the next time to use the tips with filter or you make a sterilization of the pipette by autoclave.
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