Multi-Media Direct Culture Method with DNA Fluorochrome Staining Assay

M-250 Procedural Summary

This test consists of a multi-media Direct Culture method using three (3) different mycoplasma media formulations combined with an Indicator Cell Culture procedure. The Indicator Cell Culture procedure with DNA fluorochrome assay enhances the detection of non-cultivable Mycoplasma species. This is the OPTIMUM test for detecting Mycoplasma contamination in cell cultures. (GMP service)

Sample Preperation: 

 

ADHERENT Cells:

Grow adherent cell culture in a T-25 flask. We recommend the sample have undergone at least three passages in antibiotic free media to achieve the highest level of assay sensitivity. When the cell culture is nearly confluent, scrape and aliquot 5 mL of the suspension. Dispense sample in sterile screw cap 15 mL polypropylene centrifuge tube for shipment under frozen or ambient conditions. Alternatively, sample may be submitted in a T-25 flask ambient. In this situation, simply add fresh antibiotic free media to the vessel prior to shipping. Leave sufficient room in the neck of the flask to allow for expansion and to prevent leakage. It is recommended that a back-up sample be retained and stored at -60°C or colder.

NONADHERENT Cells:
Take 5 mL of non-adherent cell suspension and place in a sterile screw cap 15 mL polypropylene centrifuge tube. We recommend the sample has undergone at least three passages in antibiotic free media to achieve the highest level of assay sensitivity. Cell culture samples can also be received in a T-25 flask ambient. In this situation, simply add fresh antibiotic free media to the vessel prior to shipping. Leave sufficient room in the neck of the flask to allow for expansion and to prevent leakage. It is recommended that a back-up sample be retained and stored at ≤ -60°C.

NOTE: Samples submitted in cryovials will be assessed a 20% surcharge for additional sample preparation.

 

Shipping Instructions:

Ship frozen on dry ice or under ambient conditions. Bionique recommends frozen shipment on dry ice. Ship samples overnight in an insulated container with sufficient dry ice to keep frozen for a period of at least 48 hours. For ambient temperature transport, place sample vessel inside a plastic bag in a crush proof package and ship overnight. Samples should be well insulated from temperature extremes.

Please view our general sample submission instructions page for details on how to submit your sample(s) online.



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GMP Service
Total Testing Time: 28 days
Sample Requirements: Minimum Test Volume = 5 mL

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