Helicobacter pylori (Marshall et al.) Goodwin et al.

1. Open thawed vial according to enclosed instructions or visit www.atcc.org for instructions.
2. Aseptically transfer the entire contents to a 5-6 mL tube of #18 broth. Additional test tubes can be inoculated by transferring 0.5 mL of the primary broth tube to these secondary broth tubes.
3. Use several drops of the primary broth tube to inoculate a #260 plate and/or #260 agar slant.
4. Or, to obtain a biphasic culture, add several drops of the primary broth tube to a #260 agar slant. Best practice is to incubate these slants at an angle.
5. Incubate at 37°C under microaerophilic conditions for 48 hours. Use an anaerobe jar with an active catalyst and a microaerophilic gas generator pack or other acceptable method. All tubes and slants should be incubated with caps loosened.
6. If growth is observed only in the primary broth, re-inoculate secondary tubes and plates from the primary broth after it shows growth.  Transfers should grow well once this initial growth is achieved.

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