Mycoplasma Detection Kit was designed to specifically detect potential Mycoplasma contamination in cell cultures. The kit incorporates polymerase chain reaction (PCR) to amplify the conserved 16S ribosomal RNA coding region within the Mycoplasma genome, thereby providing an extensive, highly sensitive and efficient detection method. Primer sequences were carefully determined to cover three genera of Mycoplasmatales (Mycoplasma, Acholeplasma and Ureaplasma) which allows this kit to detect over 95% of potential cell culture infections. The PCR technology in the Mycoplasma Detection Kit is fast (results are typically obtained in less than 3 hours) and easy to use. The kit is also highly sensitive and can detect as little as 2-5 femtograms of Mycoplasma DNA in 100uL of test sample supernatant. Eukaryotic and bacterial DNA from cell culture supernatant is not amplified by the kit. A test cell line infected with Mycoplasma will generate a PCR product(s) between ~448-611- bp on an agarose gel, depending on the type of Mycoplasma present. A positive control (M. orale, 503-bp) has been included to validate that the PCR amplification process has occurred as well as to confirm the size of the PCR product obtained in test samples. An internal control is also provided to eliminate potential false negatives associated with PCR inhibitors.