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Culture Medium and Culture Conditions
Media
LB Medium
- weight components
- 5 g/L NaCl
- 10 g/L tryptone
- 5 g/L yeast extract
- (15 g/L agar for plates)
- fill up to 1 L with deionized water
- mix well by shaking
- autoclave
- autoclaving tape, caps slightly unscrewed
- base of the pot has to be covered with deionized water
- close lid
- heat level 3 until the pressure valve opens
- reduce heat level to 1.5
- set timer to 20 minutes
- turn heater off
- wait until the pressure valve retracts (30-45 minutes)
- open, close caps & shake
- for plates, wait until you can touch the bottle (<60 °C, clean bench!)
- add antibiotics (1 µL/mL) and shake (gloves!)
TB Medium
- components 1:
- 4 mL/L glycerol
- 12 g/L tryptone
- 24 g/L yeast extract
- fill up to 900 mL with deionized water
- mix well by shaking
- autoclave
- components 2:
- 0.17 M KH2PO4
- 0.72 M K2HPO4
- dissolve in 100 mL deionized water and sterilize it by passing it through a filter
- after autoclaving and cooling down, add sterile phosphate solutions
Hartmans Minimal Medium (HM)
M9 Minimal Medium (M9)
Components for 1 L | Volume
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bidest. water | 778.667 mL
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10x Salt solution | 100 mL
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Magnesiumsulfatehaptahydrate (10 mM) | 100 mL
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Glucose 20% (w/v) | 20 mL
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1000x Trace elements | 1 mL
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Thiamin (1 mM) | 0.333 mL
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10x Salt solution
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Component' | Final concentration | Concentration in stock solution
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BisTris | 95 mM | 200,000 mg/L
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Ammmonium chloride | 60 mM | 32,100 mg/L
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Sodium citrate | 12.5 mM | 27,000 mg/L
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Monopotassium phosphate | 3 mM | 4,170 mg/L
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Dipotassium phosphate | 0.7 mM | 1,590 mg/L
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1000x Trace elements
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Component | Final concentration | Concentration in stock solution
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Iron(III) chloride | 50 mM | 13,515 mg/L
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Calcium chloride | 20 mM | 2,220 mg/L
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Manganese(II) chloride | 10 mM | 1,258 mg/L
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Zinc sulfate | 10 mM | 1,615 mg/L
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Cobalt(II) chloride | 2 mM | 260 mg/L
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Copper(II) chloride | 2 mM | 269 mg/L
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Nickel(II) chloride | 2 mM | 259 mg/L
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Sodium molybdate | 2 mM | 412 mg/L
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Sodium selenite | 2 mM | 346 mg/L
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Boric acid | 2 mM | 124 mg/L
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Hydrochloric acid | 1 mM | 20 mL
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SOC
- components
- 0,5 % yeast extract
- 2 % tryptone
- 10 mM NaCl
- 2.5 mM KCl
- 20 mM MgSO4
- fill up with deionized water
- adjust to pH 7.5 with NaOH
- after autoclaving, add 20 mM sterile glucose solution (filter sterilization)
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