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PhoenixDx® Mycoplasma Mix

Save your Precious Cell Culture!

Mycoplasma Contamination in Cell Culture

Contamination with mycoplasma is amongst the most notorious issues associated with cell cultivation. Depending on cell type, source and culture methods, the contamination with mycoplasma, acholeplasma and ureaplasma varies between 15 and 80%1. A contamination is not only inconvenient, but also a costly issue as it often requires the elimination of precious cultures.

Furthermore, the clinical use of cultured cells makes testing for mollicutes a necessity, especially for pathogens like M. pneumoniae, M. genitalium and M. hominis. PhoenixDx® offers a fast, sensitive and highly specific detection system for more than 130 mollicute species to provide certainty when certainty is needed.

qPCR for fast, sensitive and specific detection of mollicutes

Especially when working with virus samples in the cell culture, qPCR is superior to conventional detection methods as high concentrations of enveloped viruses can bias colorimetric methods like ATP conversion2.

With PhoenixDx® Mycoplasma Mix, the 16s rDNA of more than 130 mollicute species is targeted covering Mycoplasma, Acholeplasma and even Ureaplasma, whereas genomic, eukaryotic DNA (e.g. from the cell culture) is not amplified.

To exclude false-negatives due to PCR inhibition, an additional PCR positive control is included in the mastermix. PhoenixDx® Mycoplasma Mix is not only highly specific, but also easy and fast to use due to its convenient 2X mastermix formulation.

The PhoenixDx® Mycoplasma Mix includes:

  • Primers for the amplification of the 16s rDNA region
  • A PCR Positive control
  • FAM Probe for the detection of mollicutes
  • HEX Probe for the detection of the PCR Positive Control
  • dUTP for optional UNG treatment
  • Taq-antibody and VitaTaq® Polymerase for High Performance and HotStart Control
  • Optimized buffer system for an efficient lysis of intact mollicute cells
  • Universal ROX concentration for maximum instrument compatibility

Application Data of PhoenixDx® Mycoplasma Kit

PhoenixDx® Mycoplasma Mix was extensively tested for specificity against 45 non-mycoplasma DNAs. The specificity test covered viral DNA, bacterial DNA, fungal DNA and human DNA to exclude cross-reactivity with the human DNA present in cell culture environment.

Linearity & Sensitivity

PhoenixDx® Mycoplasma Mix was evaluated for linearity and sensitivity using a dilution series of mycoplasma DNA from 100 000 copies to 1 copy.

PhoenixDx® Mycoplasma Mix exhibits linearity over a broad range of target DNA input

Down to 8 copies of a mycoplasma genome can be detected.

PhoenixDx® Mycoplasma Mix…

  • is more sensitive than staining methods (detection of less than 10 genomes /reaction)
  • is specific (conserved 16s rDNA target for mollicutes)
  • is reliable (internal PCR positive control)
  • has less false-positive results than enzyme-based methods
  • has less false-Negative results than enzyme-based methods
  • requires short hands-on time and results can be obtained in less than 2 hours

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