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Contents 1 pH changing 2 Preparing medium for natto 3 Extreme Overnight cultures for Gibson 4 Gibson for rocF

pH changing

We brought hepes (which is similar to homopipes) to pH 7 then autoclaved it.

Preparing medium for natto

We prepared antibitoic ,medium for natto: 17.5g of broth in 1L, so 1.75 in 100ml

Extreme Overnight cultures for Gibson

1. G1 (T1) 1 Colony 1,
2. G1 (T2) 7 Colonies 2-6
3. G2 (T3) 2 Colonies 9,10
4. G2 (T4) 14 Colonies 11-24
5. G3 (T5) 3 Colonies 25-27
6. G3 (T6) 2 Colonies 28-29
7. G4 (T7) 4 Colonies 30-33
8. G4 (T8) 9 Colonies 34-42
9. G5 (T9) 0
10. G5 (T10) 2 Colonies 43-44

Gibson for rocF

• Promoter 136bp - 0.7 microl
• rocF 1 246bp - 1.2 microl
• rocF 2 597bp - 1.9 microl
• rocF 3 125bp - 0.4 microl
• DTERM 116bp - 0.8microl

Total: 1190bp