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Mycoplasma contamination is a common problem with continuously cultured cell lines. Contamination, which may go undetected for long periods of time, can alter cellular proliferation, gene expression, and other cellular responses. Early detection of this insidious pest is an important part of good tissue culture quality control. Here we describe the detection of Mycoplasma using the Synergy™ 2 Multi-Detection Microplate Reader to run a MycoAlert® Mycoplasma detection kit from Cambrex.
Mycoplasma are primitive prokaryotes, lacking a cell wall and consisting of only a plasma membrane, ribosomes, and a double stranded 580 kb genome. Due to their small size, mycoplasma can pass through 0.22 and 0.45 mm filters typically used to sterilize tissue culture reagents. Nor can they be observed using routine microscopy.
Mycoplasma are often slow-growing contaminants that typically exist in low numbers, but can cause experimental artifacts in cells. Mycoplasma contamination has been shown to alter cellular growth rates, affect amino acid and nucleic acid metabolism, affect cell antigenicity, and induce chromosomal and membrane aberrations.
Detection of mycoplasma can be accomplished by either direct culture techniques of the organism or by indirect assay methods. Indirect methods, which include DNA staining, biochemical detection, nucleic acid hybridization, immunoassays, and polymerase chain reaction (PCR), have the advantage of detecting “noncultivatable” species.
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