Optimized PCR-based Detection of Mycoplasma

Why is mycoplasma contamination a serious problem for cell cultures?

The maintenance of contamination-free cell lines is essential to cell-based research. Among the biggest contaminant concerns are mycoplasma contamination. Although mycoplasma do not usually kill contaminated cells, they are difficult to detect and can cause a variety of effects on cultured cells, including altered metabolism, slowed proliferation and chromosomal aberrations. In short, mycoplasma contamination compromises the value of those cell lines in providing accurate data for life science research.

Common sources of mycoplasma contamination of cell cultures, and tips for prevention

The sources of mycoplasma contamination in the laboratory are very challenging to completely control. As certain mycoplasma species are found on human skin, they can be introduced through poor aseptic technique. Additionally, they can come from contaminated supplements such as fetal bovine serum, and most importantly from other contaminated cell cultures. Once mycoplasma contaminates a culture, it can quickly spread to contaminate other areas of the lab. Strict adherence to good laboratory practices such as good aseptic technique are key, and routine testing for mycoplasma is highly recommended for successful control of mycoplasma contamination.

Video tutorial for mycoplasma PCR-based testing kit (MP0035)

PCR-based detection of mycoplasma has become a very popular method for routine cell line maintenance, and is typically the method employed by cell repositories that offer mycoplasma detection services, as well as for protecting their own stocks. PCR-based detection methods are highly sensitive and can provide rapid results, which allows researchers to respond quickly to isolate and eliminate contamination once it is detected in comparison to the time required using microbiological techniques. The LookOut Mycoplasma PCR Detection Kit is highly sensitive, with a detection limit of only 2 genomes per μL.

Which PCR reagents for mycoplasma are the most sensitive and specific?

Specific results from PCR screening for mycoplasma can be enhanced with careful reagent selection. JumpStart Taq DNA Polymerase minimizes non-specific sequence amplification and maximizes yield, enhancing both specificity and sensitivity. The LookOut Mycoplasma PCR kit also uses a proprietary primer design, significantly reducing the potential for false positive results. The 8-tube format and strips pre-coated with dNTPs helps increase the throughput to meet the needs of customers with larger collections of cell lines.

Due to the extreme sensitivity of the kit, strict aseptic technique should be followed to prevent inadvertent contamination of samples and reagents. The step-by-step video protocol clearly demonstrates necessary precautions and best practices required for reliable mycoplasma detection at a fraction of the cost of sending samples out for testing. To ensure success with the method, the video protocol includes typical results and discusses their interpretation.