Contributors: Tank, Chester
Repeated the growth-condition setup from 11–12 August (culture 3’s recipe), but substituting cellobiose for glucose in half the cultures, and comparing TOP10+beta-glucosidase against TOP10+CxnA rather than testing a single strain.
Supplement changes based on the 11–12 August results: FeSO₄ and thiamine were dropped (conditions 5 and 6 showed no significant importance for growth). Casamino acids were kept — condition 7 (no casamino acids) showed near-zero growth, confirming it’s essential.
| # | Condition | Purpose |
|---|---|---|
| 1 | M9 + glucose, full supplement (no FeSO₄/thiamine) | Main experiment |
| 2 | M9 + cellobiose, full supplement (no FeSO₄/thiamine) | Main experiment |
Chloramphenicol at 40 µg/mL, IPTG at 0.38 mg/mL throughout. Cells were washed into M9 salts using the same protocol as 11 August (OD600 of washed suspension = 2.5; inoculum V = 1000 × target-OD ÷ OD600 for 20 mL at starting OD 0.05 ≈ 400 µl).
Eight 24 h cultures set up (duplicate a/b for each strain × carbon-source combination):
- 1a/1b — glucose + beta-glucosidase
- 2a/2b — glucose + CxnA
- 3a/3b — cellobiose + CxnA
- 4a/4b — cellobiose + beta-glucosidase
No intermediate growth curve was taken this round due to time constraints — cultures were simply grown for 24 h and read the next day.