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Methanothermobacter thermautotrophicus (Zeikus and Wolfe) Wasserfallen et al.

43846

Product category
Archaea
Product type
Anaerobe
Extremophile
Strain designation
RTAE-1
Type strain
No
Isolation source
Mutant derived from strain Marburg, DSM 2133
Product format
Freeze-dried
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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

Specific applications
Biotechnology
Preceptrol
No

Characteristics

Extremophile type
Methanogen
Thermophile
Comments
5-fluorouracil-resistant

Handling information

Medium
Temperature
60°C
Atmosphere
Anaerobic
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.

3. If the medium is pink (see discussion about resazurin) add 2.0 ml of reducing agent (3% cysteine, stock solution) per 100 ml of medium.  Let the medium sit at room temperature for 10 to 20 minutes, until the resazurin becomes colorless, before inoculating.

4. After the Balch tube is ready to be inoculated let the frozen vial thaw at room temperature under a gentle stream of sterile oxygen free gas.

5. Using a 1.0 ml syringe tipped with 22 gauge needle, withdraw the cell suspension from the vial and transfer it to the broth and incubate at 37oC.  Plate 0.1 ml of the inoculated culture onto a non-selective medium and incubate aerobically at 37oC. 

6. Growth should be detected in the broth within 48 to 96 hours.  No growth should be detected on the aerobic plate or broth.

ANAEROBIC CONDITIONS:

·   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.

·   To obtain a fully reduced medium, it is necessary that the medium be anoxic and that a reducing agent be added.  Common reducing agents are sodium sulfide, cysteine, dithiothreitol, and titanium citrate.

·   Syringes can be made anaerobic by one of two methods. 

          1.     Displace the dead space in the syringe with a sterile

Handling notes
          It has been our experience the greater the headspace the better the growth.  Therefore, 25 ml Balch (Hungate) tubes with 4-5 ml of media should be used.

          It is also recommended that you pressurize the headspace with at least 20 psi to allow for good gas exchange.  For good growth, gas the culture every day for 5 to 6 days.

          Growth is noted by turbidity in the broth.  Growth is observed after approximately 48 hours.

History

Deposited as
Methanobacterium thermoautotrophicum Zeikus and Wolfe
Depositors
DP Nagle

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

Curated Citations

Nagle DP Jr., et al. 5-Fluorouracil-resistant strain of Methanobacterium thermoautotrophicum. J. Bacteriol. 169: 4119-4123, 1987. PubMed: 3624203

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