Materials and methods for mycoplasma detection and elimination

The maintenance of contamination-free cell cultures is fundamental to cell-based research. Among the most pervasive concerns is contamination with mycoplasma, a genus of bacteria whose members are typically sub-micron in size and lacking a cell wall, making them difficult or impossible to detect by microscopy and resistant to many antibiotic treatments. Although mycoplasma do not usually cause cell death, they can cause a variety of effects on cultured cells that includes altered metabolism, slowed proliferation, and chromosomal aberrations. In short, mycoplasma contamination compromises the validity of affected cell lines for providing meaningful data for life science research.

Common sources of mycoplasma in the lab, and how to control them

Sources of mycoplasma contamination in the laboratory are very challenging to control with certainty. As certain mycoplasma species are found on human skin, they can be introduced to cultures through poor aseptic technique. Additionally, mycoplasma may be introduced via contaminated supplements such as fetal bovine serum, or most commonly, by transmission from other contaminated cell cultures. Once mycoplasma is present in cell cultures, it can quickly spread to contaminate other areas of the lab in aerosols and particulates generated during culture handling. Strict adherence to good laboratory practices such as good aseptic techniques is key, and routine testing of cultures for mycoplasma is highly recommended for successful control of mycoplasma contamination.

Mycoplasma detection methods

The products below represent a complete toolkit for mycoplasma contamination detection and elimination. The three most popular mycoplasma detection methods are

  • Mycoplasma culture, where bacterial culture media optimized for mycoplasma (see table below) are inoculated with culture samples
  • DNA staining method, which is effective because mycoplasma nuclei counterstained with Hoechst or DAPI are visible by fluorescence microscopy
  • PCR (polymerase chain reaction), which will amplify the bacterial DNA if mycoplasma are present in culture samples

Mycoplasma elimination kits are a convenient, efficient choice for ridding cell cultures of mycoplasma contamination.

Materials for detection and elimination of mycoplasma contamination

Mycoplasma Detection by PCR and qPCR
Mycoplasma Detection by Mycoplasma Culture
Mycoplasma Detection by DNA Staining Method
Mycoplasma Elimination