How to Prepare LB Broth (Luria-Bertani Medium)
Standard bacterial growth medium for E. coli and other gram-negative bacteria
Components
| Chemical | Concentration |
|---|---|
| Tryptone | 10 g/L |
| Yeast extract | 5 g/L |
| NaCl | 10 g/L |
Preparation Strategy
Method: direct_dissolution
Reason: All components are >1g, suitable for direct dissolution
Preparation Table
| Component | Conc. | Amount | Unit |
|---|---|---|---|
| Tryptone | 10 g/L | 10.0 | g |
| Yeast extract | 5 g/L | 5.0 | g |
| NaCl | 10 g/L | 10.0 | g |
Procedure
- 1Step 1: Add 800 mL distilled water to a 1L beaker or flask
- 2Step 2: Add 10.0 g tryptone and mix until dissolved
- 3Step 3: Add 5.0 g yeast extract and mix until dissolved
- 4Step 4: Add 10.0 g NaCl and mix until dissolved
- 5Step 5: Adjust final volume to 1000 mL with distilled water
- 6Step 6: Mix thoroughly to ensure complete dissolution
- 7Step 7: Transfer to appropriate containers for sterilization
Sterilization
Autoclave at 121°C, 15 psi for 20 minutes. Allow to cool before use. For agar plates, add 15 g/L agar before autoclaving.
Safety
Level 1 safety: Standard laboratory gloves, lab coat, and safety glasses. No special ventilation required.
Storage
Store sterile medium at room temperature for up to 1 month or at 4°C for several months. Check for contamination before use. Prepared plates can be stored inverted at 4°C for 2-3 months.
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