Aquifex pyrophilus Huber et al.
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2. When the bottle is ready to inoculate, open the vial according to enclosed instructions.
3. For inoculation, use a 1.0 ml syringe tipped with 22 gauge needle. Withdraw 0.5 ml of 2186 and use this to rehydrate the freeze-dried pellet. Transfer the rehydrated cell into the 2186 broth; from this bottle prepare a 2nd bottle at a 1:10 dilution. Incubate the bottles at 75 - 85oC. 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 and incubate at 70oC
4. Growth should be detected in the #2186 broth within 24 - 48 hours. There should be no growth detected on the aerobic plate. There should be no growth in the nonselective aerobic or anaerobic broth.
It is important to keep the headspace-liquid ratio high as this enhances H2 and O2 transfer to the culture.
Cells appear as rods and may be motile for a short period at room temperature.
The cryoprotectant used for lyophilization may be somewhat inhibitory for growth, which is why we recommend diluting from the original inoculum.
The gas mix is flammable, use caution with the gas stream around open flames.
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