Bacterial Growth Media
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LB Broth
To 200 mL ddH2O add:
- Pre-made:
- 6.25g Miller LB powder
- 6.25g Miller LB powder
- OR
- 2.5g Tryptone
- 2.5g NaCl
- 1.25g Yeast extract
- pH to 7.5 if necessary
- 2.5g Tryptone
- Bring to 250 mL volume
Aliquot into bottles (~50 mL per bottle) for autoclaving. The bottle caps should be held very loosely, covered in foil and labelled. The bottles should be placed in a steel basket.
SOC Medium
Used to improve yield following transformation, during recovery phase.
To 35 mL ddH2O add:
- 0.375g Yeast Extract
- 1.5g Tryptone
- 0.045g NaCl
- 0.09g MgSO4
- 0.014g KCl
Bring to 75 mL volume and autoclave.
Then add:
- 0.75mL 1M MgCl2
- 0.24g Glucose
Finally, sterilize the media by passing it through a 0.2μm filter
Terrific Broth (TB)
Used to grow E. coli ZYCY10P3S2T during minicircle generation protocol
To 150 mL ddH2O in a 500 mL bottle, add:
- 4.8g Yeast Extract
- 4g Tryptone
- 4mL Glycerol
Bring to volume of 180 mL and autoclave
To 20 mL ddH2O, add:
- 0.046g KH2PO4 (potassium phosphate monobasic)
- 0.25g K2HPO4 (potassium phosphate dibasic)
Filter sterilize and add to the autoclaved portion, for final volume of 200 mL