Protocol: Mycoplasma Removal Agent (MRA)

Overview

1. Add Mycoplasma Removal Agent to cell cultures contaminated by mycoplasma at the recommended concentration of 0.5 µg/ml and incubate for 1 week
2. Mycoplasma Removal Agent concentration should be raised up to 1 µg/ml only if the recommended concentration is ineffective in removing the mycoplasma
3. For media replacement or culture transfer (passage), use a medium containing Mycoplasma Removal Agent at this same concentration
4. Transfer the cell cultures several times without Mycoplasma Removal Agent and confirm that regrowth of the contaminating mycoplasma has not occurred

Example Data

The following tables show that the level of infection, cell type and mycoplasma strains may influence specific results. We encourage researchers to use this data as a helpful guide to indicate the effective Mycoplasma Removal Agent concentration needed with their specific cell line and mycoplasma strain.

(+) = Mycoplasma positive, (-) = Mycoplasma negative, (Days*) = Mycoplasma status at stage of culture (days).

Comparisons with other antibiotics on the market such as Tiamulin and Minocycline shows that Mycoplasma Removal Agent is comparable for its minimum inhibitory concentration (MIC), with MICs from as low as 0.1 µg/ml for all the tested bacteria (see Table 2). This shows that if you are looking to inhibit the growth of Mycobacteria within your cultures, you can reliably use Mycoplasma Removal Agent because it is just as effective as other antibiotics which target mycobacteria.

Where Mycoplasma Removal Agent out performs the other tested antibiotics is in its low Minimum Microbicidal Concentration (MMC) which shows consistently low concentrations required for microbicidal effect (see Table 2). This enables you to confidently kill and remove mycoplasma from cultures at low concentrations of antibiotic. This reduces the chance that the antibiotic will have any inhibitory effects upon other cells in your culture.

*MIC = Minimum inhibitory concentration (µg/ml), **MMC = Minimum microbicidal concentration (µg/ml)

Reference

• Nakai, N. et al. (2000). Detection and elimination of contaminating microorganisms in Transplantable Tumors and Cell Lines. Exp. Anim. 49: 309 – 313.

Related Topics

• Cell Proliferation Reagents
• Antibody Manufacturing Tools
• Endotoxin Removal Kit

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