Bacterial Growth Media

LB Broth

To 200 mL ddH₂O add:

• Pre-made:
  • 6.25g Miller LB powder
    
• OR
  
  • 2.5g Tryptone
    
  • 2.5g NaCl
    
  • 1.25g Yeast extract
    
  • pH to 7.5 if necessary

• 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.

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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

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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