Culture medium and culture conditions

Media

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
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
for plates, wait until you can touch the bottle (<60 °C, clean bench!)
add antibiotics (1 µL/mL) and shake (gloves!)

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

Components for 1 L	Volume
bidest. water	778.667 mL
10x Salt solution	100 mL
Magnesiumsulfatehaptahydrate (10 mM)	100 mL
Glucose 20% (w/v)	20 mL
1000x Trace elements	1 mL
Thiamin (1 mM)	0.333 mL

Component'	Final concentration	Concentration in stock solution
10x Salt solution
BisTris	95 mM	200,000 mg/L
Ammmonium chloride	60 mM	32,100 mg/L
Sodium citrate	12.5 mM	27,000 mg/L
Monopotassium phosphate	3 mM	4,170 mg/L
Dipotassium phosphate	0.7 mM	1,590 mg/L

Component	Final concentration	Concentration in stock solution
1000x Trace elements
Iron(III) chloride	50 mM	13,515 mg/L
Calcium chloride	20 mM	2,220 mg/L
Manganese(II) chloride	10 mM	1,258 mg/L
Zinc sulfate	10 mM	1,615 mg/L
Cobalt(II) chloride	2 mM	260 mg/L
Copper(II) chloride	2 mM	269 mg/L
Nickel(II) chloride	2 mM	259 mg/L
Sodium molybdate	2 mM	412 mg/L
Sodium selenite	2 mM	346 mg/L
Boric acid	2 mM	124 mg/L
Hydrochloric acid	1 mM	20 mL