Bacterial Growth Media

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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.

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