Table 1. Kit Components and Storage

Product Description

Mycoplasma infections are relatively common in laboratory cell cultures; it has been estimated that between 5% and 35% of all cell cultures are infected. Mycoplasmas have been shown to alter the growth rate of cells in culture, induce chromosomal aberrations, influence amino acid and nucleic acid metabolism and cause membrane aberrations. Several methods have been developed to detect mycoplasma including direct culture in special growth media, enzyme-linked immunoassay, immunofluorescence staining, PCR, biochemical detection and fluorescent nucleic acid stains.

The Mycoplasma Stain Kit provides an ultrasensitive, rapid and simple fluorescence microscopic assay for the visual identification of mycoplasma infection in laboratory cell cultures. In order to detect mycoplasma, the fluorescent MycoFluor reagent is added directly to the culture medium, with or without cells, and the stained sample is then examined under a fluorescence microscope. The excitation and emission spectra of MycoFluor reagent bound to dsDNA are similar to dsDNA bound to DAPI, with excitation maxima around 350-360nm and emission maxima around 450-460nm. MycoFluor reagent can be excited either with a xenon mercury arc lamp or a UV laser and is detected through a blue filter.

Mycoplasma staining with MycoFluor reagent appears as a fine particulate or filamentous staining over the cytoplasm at 100X magnification. Nuclei of the cells are also brightly stained by this method and thereby act as endogenous positive control for the staining procedure.

Materials required but not provided in the kit:

· • Sterile tissue culture grade water

· • Sterile PBS buffer

· • Carnoy's fixative (3:1 Methanol: Glacial acetic acid)

· • Centrifuge tubes

· • Glass slides

· • Microscope coverslips