Mycoplasma Detection

Mycoplasma Detection using EZ-PCR Mycoplasma Test Kit

A highly sensitive PCR assay in kit format
Detects all Mycoplasma species and achoplasma and spiroplasma species
Quick, simple, reliable and cost-effective method for regularly monitoring cells for mycoplasma detection
Composed of a single tube PCR mix, buffer solution and positive control
The time taken for preparing the sample for PCR is less than 10 minutes

The EZ-PCR Mycoplasma Test Kit is a highly sensitive PCR assay which detects all Mycoplasma species that may contaminate tissue culture samples as well as achoplasma and spiroplasma species. It is a quick, simple and reliable and cost-effective method for regularly monitoring cells for mycoplasma detection.

EZ-PCR Mycoplasma Test Kit is composed of a single tube PCR mix. It does not require special equipment and can be performed in any lab with a thermal cycler and electrophoresis equipment.

Running the PCR reaction products on an agarose gel will result in a 270bp band.

All of the ingredients needed for the PCR step are in a pre-mixed reaction solution so there is no need to worry about buffer, enzyme and dNTP concentrations. The primers used in the kit are derived from a highly conserved region within the 16S rRNA gene and can detect very low levels of contamination.

Samples are centrifuged in a microcentrifuge tube. The pellets obtained are suspended in a unique DNA-release buffer and the DNA is amplified by PCR.

The time taken for preparing the sample for PCR is less than 10 minutes.

The kit contains Buffer Solution, PCR mix and a Positive Control sufficient for 10 or 20 tests.

Mycoplasma Prevention and Removal Agents

Geneflow now supply a complete and co-ordinated package of high quality reagents suited for dealing with all aspects of Mycoplasma control within the tissue culture laboratory.

Mycoplasma Prevention

1) Pharmacidal Disinfectant Spray

For use on all surfaces and incubators
Effective against the whole spectrum of bacteria (and spores), mycoplasma, yeast, fungi (and spores) and viruses (HBV, HIV)
Active ingredients are quarternary benzylammonium compounds
Non-toxic, does not contain mercury, formaldehyde, phenol or alcohol
Simply spray onto surface before use and allow to air-dry (aprox 15 minutes)

Pharmacidal is a ready-for-use disinfectant spray suitable for use in all areas of the laboratory. It is effective against the whole spectrum of bacteria, mycoplasma, yeast, fungi and viruses. It contains no aldehydes, phenolic derivatives, chlorine or peroxides but is based on an inventive combination of three benzyl ammonium chlorides in a specific ratio.

Based on the manufacturers experience, Pharmacidal can be used in loaded incubators without problems since the active substance is not volatile. Pharmacidal is bio-degradable and causes no damage to incubators, hoods or workers.

Tables and hoods that are used by more than 1 person should be sprayed before each person starts working instead of the common practice of illuminating UV lamps for 20 minutes prior to working.

Composition

100g Pharmacidal contains:

0.0975g n-Octyl-dimethyl-Benzylammonium-acetate
0.0300g Benzelthoniumacetate
0.0025g Methylbenzenethoniumacetate

Appearance

Clear, Colourless Solution
pH 8.0
Stability 5 Years

Microbiology

Bactericidal
Fungicidal
Virus Inactivating (HBV, HIV, Rotaviruses)

Surface Disinfection

The surfaces to be disinfected should be completely wet by spraying (Drying Time = Reaction Time). Allow the surface to dry, no rinsing is necessary apart from surfaces which come into contact with food.

2) Aquaguard Disinfectants

For adding to water in incubators and water baths
Aquaguard -1 for water disinfection in humidified CO2 incubators. It contains a disinfectant that does not cause damage to the stainless steel tray, is non-toxic and non-volatile. Use at 10ml/litre water.
Aquaguard-2 is designed for use with standard water baths and prevents bacterial and fungal growth. Use a 2ml/litre water

A CO2 incubator and water bath are basic equipment required in tissue culture. However these also make fertile breeding ground for contaminants due to the high humidity and temperature. As soon as contamination appears it can be difficult to eliminate and severe damage can be caused. It is therefore essential to systematically clean and disinfect the equipment.

Disinfecting the Water in the Incubator

Aquaguard -1 contains a disinfectant that does not cause damage to the stainless steel tray, is non-toxic and non-volatile. The water should be replaced with sterile water every two to four weeks, adding 50ml of Aquaguard-1 per 5 litres of water.

Mycoplasma Removal

Mycoplasma Removal using BIOMYC antibiotic solutions

A combination of antibiotics which have been shown to be effective in the elimination of mycoplasma species that account for 90% of the contamination found in cell cultures.
When used according to the instructions, no cytotoxic effects will occur.
BIOMYC-1 is based on the antibiotic tiamutin,
BIOMYC-2 is based on minocycline, which is a tetracycline derivative.
BIOMYC -3 is based on ciprofloxacin which is a member of the fluoroquinolone group

BIOMYC solutions are a combination of antibiotics which have been shown to be effective in the elimination of mycoplasma species that account for 90% of the contamination found in cell cultures. When used according to the instructions, no cytotoxic effects will occur.

BIOMYC-1 is based on the antibiotic tiamutin, which is produced by the fungus pleurotus mutilus. BIOMYC-2 is based on minocycline, which is a tetracycline derivative. It is recommended that BIOMYC 1 and 2 are used sequentially rather than together.

BIOMYC -3 is based on ciprofloxacin which is a member of the fluoroquinolone group. Many mycoplasma species have been found to be sensitive to BIOMYC-3 which when used at concentrations recommended for use should not demonstrate any cytotoxic effects.