Escherichia coli (Migula) Castellani and Chalmers
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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. From a single tube of #1622 broth (5 to 6 ml), withdraw approximately 0.5 to 1.0 ml with a Pasteur or 1.0 ml pipette and use to rehydrate the pellet.
3. Aseptically transfer the rehydrated pellet back into the broth tube. Mix well.
4. Use several drops of this suspension to inoculate a second tube of broth, a slant, and/or a plate.
5. Incubate all tubes and plate at 37°C for 24 hours.
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Schellhorn HE, Hassan HM. Isolation and characterization of respiratory-deficient mutants of Escherichia coli K-12. J. Bacteriol. 170: 78-83, 1988. PubMed: 3275633