Wittrup: E. coli Media

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LB Media [see also LB]

Liquid, 1000 mL

  • 10g Bacto tryptone
  • 5g Bacto yeast extract
  • 10g NaCl

pH to 7.5 with NaOH (optional?) Add ddH2O to final volume. Filter sterilize, or autoclave. Add antibiotic if selective media is desired (perhaps 100 mg/L amp), but do not add until cooled to below 50 oC (touchable by bare hand)

Plates, 1000 mL

  • Ingredients as above (still save antibiotic for last)
  • 15-18g agar
  • Autoclave. Add antibiotic if selective media is desired, but do not add until cooled to below 50 oC (touchable by bare hand)

Ampicillin [see also Ampicillin]

  • Stock: 25 mg/mL of sodium salt of ampicillin (amp) in water.
  • Filter sterilize and store in aliquots of 4 mL at -20 oC
  • Working concentration = 35-50 mg/L for liquid media. Often 100 mg/L for plates.

SOC Media [see also SOC]

100 mL

  • 2.0g 2% Bactotryptone
  • 0.5g 0.5% Yeast extract
  • 10 mM NaCl (0.2 mL 5 M)
  • 10 mM MgCl2 (1.0 mL 1 M)
  • 10 mM MgSO4 (1.0 mL 1 M)
  • 20 mM Dextrose (D-glucose) (0.36 g)

Add ddH20 to final volume. Filter sterilize, or autoclave.

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