ATCC determines the biosafety level of a material based on our risk assessment as guided by the current edition of Biosafety in Microbiological and Biomedical Laboratories (BMBL), U.S. Department of Health and Human Services. It is your responsibility to understand the hazards associated with the material per your organization’s policies and procedures as well as any other applicable regulations as enforced by your local or national agencies.
ATCC highly recommends that appropriate personal protective equipment is always used when handling vials. For cultures that require storage in liquid nitrogen, it is important to note that some vials may leak when submersed in liquid nitrogen and will slowly fill with liquid nitrogen. Upon thawing, the conversion of the liquid nitrogen back to its gas phase may result in the vial exploding or blowing off its cap with dangerous force creating flying debris. Unless necessary, ATCC recommends that these cultures be stored in the vapor phase of liquid nitrogen rather than submersed in liquid nitrogen.
2. Exchange the gas in the test tube for 100% N2; do not over pressurize.
3. Prepare tubes for inoculation: If the medium is pink (see B), add 0.1 ml of reducing agent (5% Co-enzyme M stock solution; see C) per each 5 to 10 ml of medium. Let the medium sit at room temperature for at least 2 hours, or until the resazurin becomes colorless, before inoculating.
4. Thaw the frozen vial under a gentle stream of anaerobic gas. Using an anaerobic (see D) 1.0 ml syringe tipped with 22-gauge needle, withdraw the cell suspension from the vial and transfer to the tube of ATCC #2731 medium. Transfer 0.5 ml of the inoculated culture to a second tube of ATCC® medium #2731. Plate 0.1 ml of the inoculated culture onto a non-selective medium and incubate the plate aerobically at 37oC. Incubate culture tubes at 55oC.
5. Growth should be detected in the broth within 1 to 2 days. No growth should be detected on the aerobic plate.
Additional information on this culture is available on the ATCC® web site at www.atcc.org.
The product is provided 'AS IS' and the viability of ATCC® products is warranted for 30 days from the date of shipment, provided that the customer has stored and handled the product according to the information included on the product information sheet, website, and Certificate of Analysis. For living cultures, ATCC lists the media formulation and reagents that have been found to be effective for the product. While other unspecified media and reagents may also produce satisfactory results, a change in the ATCC and/or depositor-recommended protocols may affect the recovery, growth, and/or function of the product. If an alternative medium formulation or reagent is used, the ATCC warranty for viability is no longer valid. Except as expressly set forth herein, no other warranties of any kind are provided, express or implied, including, but not limited to, any implied warranties of merchantability, fitness for a particular purpose, manufacture according to cGMP standards, typicality, safety, accuracy, and/or noninfringement.
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