Gen5 version required: 1.0 or higher
Basis for the Assay:
Mycoplasma contamination is a common problem with continuously cultured cell lines. The MycoAlert® assay from Cambrex is a biochemical test that screens for the presence of mycoplasma-specific enzymes that interact with reagents catalyzing the conversion of ADP to ATP. By measuring the ATP produced before and after the addition of a substrate, a ratio is obtained which indicates the presence or absence of mycoplasma.
This procedure includes a 5-minute incubation in the reader, a first 1-second per well integration, then a “plate eject” command. This command allows the manual addition of the MycoAlert substrate. The plate is then incubated for ten minutes in the reader before a second read is performed. The protocol calculates the MycoAlert ratios, interprets the results (positive, negative, re-test) and plots a standard curve.
The file incorporates data from a serial dilution of a positive control.
Plate configuration includes a serial dilution of a positive control. 9 dilutions of the control are available, each dilution in replicates of 8.
The report lists the Procedure and Data Reduction steps, and includes:
1. Standard curve and curve fitting results table
2. Plate layout and interpreted results (Cutoffs) in a matrix format
3. Plate layout and calculated concentrations in a matrix format