Cultivation of Bacteria

1. Inoculate 2% of the microorganisms in 200 mL MRS broth (see Note 1).

2. Incubate overnight at 30°C (mesophilic bacteria) or 37°C (thermophilic bacteria) (see Note 2).

3. Collect cells by centrifugation at 10,000g for 10 min at 4°C.

4. Wash the pellet twice with 10 mL of 0.1 M sodium phosphate buffer, pH 7.0.

5. Resuspend at 40% w/v washed cells in the afore-mentioned buffer by vortexing. Be sure that the pellet is completely resuspended before proceeding.

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