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Methods Mol Biol. 2013;946:1-13. doi: 10.1007/978-1-62703-128-8_1.

Detection of mycoplasma contaminations.

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Department of Human and Animal Cell Lines, DSMZ, German Collection of Microorganisms and Cell Cultures, Braunschweig, Germany.


Mycoplasma contamination of cell lines is one of the major problems in cell culture technology. The -specific, sensitive, and reliable detection of mycoplasma contamination is an important part of mycoplasma control and should be an established method in every cell culture laboratory. New cell lines as well as cell lines in continuous culture must be tested in regular intervals. The polymerase chain reaction (PCR) methodology offers a fast and sensitive technique to monitor all cultures in a laboratory and can also be used to determine the contaminating species.The described assay can be performed in less than 3 hours, including sample preparation, DNA extraction, PCR run, and analysis of the PCR products. Special emphasis is given to steps taken to avoid false-negative results due to the presence of inhibitors of the Taq polymerase in the crude samples and the interpretation of the results. The technique can also be adapted to the requirements of the pharmacopoeia.

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