all the manual or tips about LAMP think that false positive will hapen if you open the PCR tube for gel electrophoresis detection. How can it been for the enzyme has been inactived by 85 degree for 10 min?
Can anyone give me some suggestions
Hi, the big problem about LAMP is not the enzyme, but the high amount of DNA you are generating, so the contamination when opening the PCR tubes comes from this DNA, you can generate aerosols and have air contamination, so you must be very careful if doing gel electrophoresis. Also, as LAMP is so sensitive, even the slightest amount DNA can lead to false positives.
The best suggestion I have is to have different rooms (One for PCR Master Mix, and one for electrophoresis), always use filter tips and use working stocks for every reagent, so if you encounter contamination, only the stocks you are using may be contaminated and not the original reagent.
Also check very carefully the primers, because it happens a lot that the primers have self amplification, so the problem doesn't come from contamination.
I hope this helps a bit.
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