43587
43587 ™
Download Genome Learn about the ATCC Genome Portal43587 ™
Download Genome Learn about the ATCC Genome PortalATCC 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. If needed exchange the gas in the test tube for 100% N2. Best results are obtained if the tube is pressurized to approximately 1.52 bar.
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. When the Balch tube is ready to inoculate, open the vial according to enclosed instructions.
5. For inoculation, use an anaerobic 1.0 ml syringe (see below) tipped with 22 gauge needle. Withdraw 0.5 ml of #1915 broth and use this to rehydrate the freeze dried pellet. Immediately place the rehydrated vial under a stream of sterile oxygen-free gas.
6. Using the same syringe, transfer the rehydrated cell suspension back to a tube of #1915 broth. Plate 0.1 ml of the inoculated culture onto a non-selective medium and incubate aerobically at 37oC. Inoculate a nonselective anaerobic and aerobic broth. Incubate the inoculated tubes at 90 to 100oC.
7. Growth should be detected in the #1915 broth within 24 hours. There should be no growth detected on the aerobic plate. There should be no growth in the nonselective aerobic or anaerobic broth.
ANAEROBIC CONDITIONS:
a. 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).
b. 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.
c. 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.
d. Syringes can be made anaerobic by one of two methods. 1. Displace the dead space in the syringe with a sterile
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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.
Fiala G, Stetter KO. Pyrococcus furiosus sp. nov. represents a novel genus of marine heterotrophic archaebacteria growing optimally at 100 celsius. Arch. Microbiol. 145: 56-61, 1986.
Validation list no. 22. Int. J. Syst. Bacteriol. 36: 573-576, 1986.
Halio SB, et al. Sequence, expression in Escherichia coli, and analysis of the gene encoding a novel intracellular protease (PfpI) from the hyperthermophilic archaeon Pyrococcus furiosus. J. Bacteriol. 178: 2605-2612, 1996. PubMed: 8626329
type strain