Mycoplasmas are the smallest and simplest prokaryotes, which often infect cell cultures in laboratories and have an effect on their physiology and metabolism. We used different tests, including PCR, DNA staining and commercial kits, to inspect our cell cultures for mycoplasma contamination. We have shown, that some of our most frequently used cell cultures are contaminated with mycoplasmas. To find out the species of contaminating mycoplasmas, we used sequencing and compared our sequence to known species. All checked cell cultures were contaminated with Mycoplasma hyorhinis. To eradicate the contamination, we added antibiotic gentamycin in high concentration directly to the cell medium, but the cells remained mycoplasma contaminated, probably due to antibiotic resistance. The Polymerase chain reaction turned out to be the most useful method for detection of mycoplasmas in cell culture because of its reliability, price, simplicity and possibility of analysing more samples at the same time.
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