Variable | Phase 0 | Phase 1 | Phase 2 |
---|---|---|---|
Cryoprotection Media | SM > 17%G:TSB | NA | NA |
Inoculum (colonies) | NA | 10 > 5 or 15 or loopful | NA |
Culture Broth | NA | THB > BHI | THB > BHI |
Yeast Extract (% YE) | 0.5 > 0 | 2.0 > 0 | 2.0 > 0 |
Horse Blood (% HB) | NA | 2.5 > 0 | 2.5 > 0 |
pH adjustment to 7.8 | NA | Initially > NoAdj | q1h > NoAdj |
Incubation time (h) | 15 > 18 > 24 | 12 > 15, 18, 24 | 3*, 5†, 6‡ |
Ideal dilution (Tube #) | NA | 5 > 4 > 1–3 | 6 = 7 = 8 |