When checking our cell cultures by electron microscopy reveals the formation like mycoplasmas. Structure is very similar to the mycoplasma. However, no staining DAPI, no matter the culture method, we do not detect mycoplasma. What could it be?
Agree with a previous comment. DAPI staining is not sensitive enough, so to be completely sure you should perform PCR
I also agree with my 2 colleagues here. I also performed DAPI staining, but it was somehow useless. A PCR check is the best way to check your cultures. There are several commercial kits available. Or just make your own primers. Does your culture grow slowly? This is also a hint for mycoplasma contaminations.
I agree with you. Recently we had a PCR test (Universal Mycoplasma Detection Kit, ATCC) and the results were negative.
As mentioned by others PCR is the most sensitive methods for mycoplasma detection. If all your controls, positive, negative and sample controls are coming up as they should and you are getting no mycoplasma coming up, you need to check the strains of mycoplasma that your primers pick up. We have "in house" primers for mycoplasma and I know it's difficult to get primers to cover all common mycoplasma strains. And yes, Dapi will not show up low level mycoplasma. You might want to consider treatment if you are especially worried. Invivogen have some good products like plasmocure that you can use.
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