New kit Ames MPF PENTA 2 is identical to Ames MPF PENTA 1, but it includes the strain E.coli WP2 uvrA(pKM101), instead of the two strains E.coli WP2 uvrA and E.coli WP2 [pkM101] resulting in the E.coli Combo. The new kit has the advantage of having only 1 E.coli strain which is listed in OECD 471. Proficiency chemicals have been tested.
For details of the Ames Penta technology and how it works please click here
Ames MPF Penta 1 (Short Protocol)
Bacteria are exposed to 6 concentrations of a test agent, as well as a positive and a negative control, for 90 minutes in medium containing sufficient histidine (S. typhimurium) or tryptophan (E.coli) to support approximately two cell divisions. After exposure, the cultures are diluted in pH indicator medium lacking histidine or tryptophan and aliquoted into 48 wells of a 384-well plate. Within two days, cells that have undergone reversion to amino acid prototrophy will grow into colonies.
Bacterial metabolism reduces the pH of the medium, changing the color of that well. The number of wells containing revertant colonies are counted for each dose and compared to a solvent (negative)control. Each dose is done in triplicate to allow for statistical analysis of the data. A dose dependent increase in the number of revertant colonies upon exposure to test sample relative to the solvent control indicates that the sample is mutagenic in the Ames MPF assay.The mutagenic potential of samples is assessed directly and in the presence of liver S9 fractions.
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