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Methanosarcina mazei (Barker) Mah and Kuhn

BAA-2030

Product category
Archaea
Product type
Extremophile
Strain designation
O1M9701 [OCM 748]
Type strain
No
Product format
Frozen
Storage conditions
-80°C or colder
Mission Collection Item
This is a Mission Collection Item.

Documentation

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.

Detailed product information

General

Preceptrol
No

Characteristics

Extremophile type
Methanogen
Methylotroph
Comments
This item is part of The Oregon Collection of Methanogens (OCM) that was kindly donated to the ATCC by Dr. David Boone. This collection is now part of the ATCC general collection and available to the scientific community. OCM items are archived as original material and available on a "made-to-order" basis. ATCC will make a reasonable effort to recover these items from preservation, but recovery and identification is not guaranteed.

Handling information

Medium
Temperature
37°C
Atmosphere
80% H2, 20% CO2
Handling procedure
  1. Sterilize the top of the Balch tube by spraying it with 70% ethanol and then flaming the top.
  2. If needed exchange the gas in the test tube for 80% H2 -20% CO2; do not go above 5 PSI.
  3. If the medium is pink (see discussion about resazurin) add  0.1 mL reducing agent (1.5% Na2S.9H2O, stock solution) per each 5-6 mL of medium.  Let the medium sit at room temperature for 10 to 20 minutes - until the resazurin becomes colorless - before inoculating.
  4. When the Balch tube is ready to inoculate, open the vial thaw in an oxygen free environment.
  5. For inoculation, use a 1.0 mL syringe tipped with 22 gauge needle.  Make the syringe anaerobic (see discussion below) and draw the thawed cell suspension up into the syringe.  Transfer the cell suspension into a tube of pre reduced #2467 broth and incubate at 30°C.  Secondary tubs can be inoculated by transferring 0.5 mL of the primary tube.  Plate 0.1 mL of the inoculated culture onto a non-selective medium and incubate aerobically at 30°C. Inoculate a nonselective anaerobic and aerobic broth and incubate at 30°C
  6. Growth should be detected in the #2467 broth within 5 to 7 days.  The gas in the head space of the Balch tube should be exchanged for fresh 80% H2 -20% CO2 every 2 to 3 days.  There should be no growth detected on the aerobic plate.  There should be no growth in the nonselective aerobic or anaerobic broth.

 

ANAEROBIC CONDITIONS:

  1. Balch tube refers to a special type of test tube that is designed to be pressurized and is suited for anaerobic work.  The Balch test tubes can be purchased form Bellco glass (www.bellcoglass.com; stock no. 2048-00150).
  2. Resazurin is a commonly used redox indicator that is pink when the redox potential is above -50 mv.,  and colorless when the redox potential is below -110 mv. i.e. highly reducing.  Most strict anaerobes require this low redox potential for optimum growth.
  3. To obtain a fully reduced medium, it is necessary that the medium be anoxic and that a reducing agent be added.
  4. Common reducing agents are sodium sulfide, cysteine, and coenzyme-M (See below).
  5. Syringes can be made anaerobic by one of two methods. 
    1. Displace the dead space in the syringe with a sterile oxygen-free gas.  
    2. Displace the dead space in the syringe with a reducing agent
Handling notes
If acetate or methanol is included in the medium then the 80% Hydrogen-20% Carbon dioxide gas does not need to be exchanged.
Additional information on this culture is available on the ATCC® web site at www.atcc.org.

History

Depositors
D Boone
Special collection
Oregon Collection of Methanogens

Legal disclaimers

Intended use
This product is intended for laboratory research use only. It is not intended for any animal or human therapeutic use, any human or animal consumption, or any diagnostic use.
Warranty

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.

Disclaimers

This product is intended for laboratory research use only. It is not intended for any animal or human therapeutic use, any human or animal consumption, or any diagnostic use. Any proposed commercial use is prohibited without a license from ATCC.

While ATCC uses reasonable efforts to include accurate and up-to-date information on this product sheet, ATCC makes no warranties or representations as to its accuracy. Citations from scientific literature and patents are provided for informational purposes only. ATCC does not warrant that such information has been confirmed to be accurate or complete and the customer bears the sole responsibility of confirming the accuracy and completeness of any such information.

This product is sent on the condition that the customer is responsible for and assumes all risk and responsibility in connection with the receipt, handling, storage, disposal, and use of the ATCC product including without limitation taking all appropriate safety and handling precautions to minimize health or environmental risk. As a condition of receiving the material, the customer agrees that any activity undertaken with the ATCC product and any progeny or modifications will be conducted in compliance with all applicable laws, regulations, and guidelines. This product is provided 'AS IS' with no representations or warranties whatsoever except as expressly set forth herein and in no event shall ATCC, its parents, subsidiaries, directors, officers, agents, employees, assigns, successors, and affiliates be liable for indirect, special, incidental, or consequential damages of any kind in connection with or arising out of the customer's use of the product. While reasonable effort is made to ensure authenticity and reliability of materials on deposit, ATCC is not liable for damages arising from the misidentification or misrepresentation of such materials.

Please see the material transfer agreement (MTA) for further details regarding the use of this product. The MTA is available at www.atcc.org.

Permits & Restrictions

Import Permit for the State of Hawaii

If shipping to the U.S. state of Hawaii, you must provide either an import permit or documentation stating that an import permit is not required. We cannot ship this item until we receive this documentation. Contact the Hawaii Department of Agriculture (HDOA), Plant Industry Division, Plant Quarantine Branch to determine if an import permit is required.

MORE INFORMATION ABOUT PERMITS AND RESTRICTIONS

Frequently Asked Questions

References

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