Culture medium and culture conditions

Media

LB medium

1. weight components

   5 g/L NaCl

   10 g/L tryptone

   5 g/L yeast extract

   (15 g/L agar for plates)

2. fill up to 1 L with deionized water
3. mix well by shaking
4. autoclave
   1. autoclaving tape, caps slightly unscrewed
   2. base of the pot has to be covered with deionized water
   3. close lid
   4. heat level 3 until the pressure valve opens
   5. reduce heat level to 1.5
   6. set timer to 20 minutes
   7. turn heater off
   8. wait until the pressure valve retracts (30-45 minutes)
   9. open, close caps & shake
5. for plates, wait until you can touch the bottle (<60 °C, clean bench!)
6. add antibiotics (1 µL/mL) and shake (gloves!)

TB medium

1. components 1:

   4 mL/L glycerol

   12 g/L tryptone

   24 g/L yeast extract

2. fill up to 900 mL with deionized water
3. mix well by shaking
4. autoclave
5. components 2:

   0.17 M KH₂PO₄

   0.72 M K₂HPO₄

6. dissolve in 100 mL deionized water and sterilize it by passing it through a filter
7. after autoclaving and cooling down, add sterile phosphate solutions

Hartmans minimal medium (HM)

M9 minimal medium (M9)

SOC

1. components

   0,5 % yeast extract

   2 % tryptone

   10 mM NaCl

   2.5 mM KCl

   20 mM MgSO₄

2. fill up with deionized water
3. adjust to pH 7.5 with NaOH
4. after autoclaving, add 20 mM sterile glucose solution (filter sterilization)