We are facing a lot of contamination problems.
Is better to check by PCR with the cell supernatant!!! When you can see it by microscopy, you have a huge infection!!! You should detect before that
Thank you. if i will add more antibiotic to the medium means it will kill the microbes or what happend to the cells?
can i use sigma products or invitrogen products? Complete medium preparation 10 mmHEPES is essential r not?. NEE also essential?
In Our Medium Composition for 1 liter ?
DMEM invitrogen
3.7g sodium bicarbonate (Himedia)
1% antibiotic&antimycotic (Sigma)
10mM HEPES (Himedia)
Mycoplasma infection can be really difficult to remove, especially if well established. Commercial mycoplasma removal agents are expensive but can still fail.
If you're working with such common lines as A549 or MCF7, it would be normal practice to ditch the infected batch and start with a fresh vial (or buy a vial of the line again if all your stock is infected).
If you absolutely need to cure your cells (e.g. they are your own clones, or something else very valuable), start off with trying ciprofloxacin treatment (protocols widely available online) for a minimum of 14 days. This is cheap and a good starting point, before trying any of the much more expensive reagents.
Finally, if you're getting regular infections in your lab, clean and decontaminate it as thoroughly as possible. You can buy cleaning solutions/sprays to get rid of myco contamination, which we've had to do for our hoods before.
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