Mycoplasma PCR controls are essential components in the detection and monitoring of Mycoplasma contamination in cell culture environments. Mycoplasmas are small, self-replicating bacteria that lack a cell wall, making them resistant to many antibiotics commonly used in cell culture. Contamination can significantly affect cellular physiology, leading to unreliable experimental results.

Content of Mycoplasma PCR Controls

Positive Control

• Contains a known quantity of Mycoplasma DNA.
• Ensures the PCR assay is functioning correctly by verifying that the PCR reagents, thermal cycling conditions, and equipment are capable of detecting Mycoplasma DNA.
• Helps to confirm the sensitivity and specificity of the assay.

Negative Control

• Contains all PCR reagents without Mycoplasma DNA.
• Validates the absence of contamination in the PCR reagents and laboratory environment.
• Critical for identifying false positives that may arise from contamination during sample preparation or PCR setup.

Internal Control

• A non-target DNA sequence added to each reaction to confirm that PCR inhibition is not present in the samples.
• Helps to differentiate between a true negative result (no Mycoplasma present) and a false negative result (Mycoplasma present, but not detected due to PCR inhibition).

Reference Control

• A well-characterized sample with a known Mycoplasma contamination level.
• Used to assess the quantitative performance of the PCR assay, such as determining the limit of detection (LOD) and ensuring consistent performance across different batches of reagents or different runs of the assay.