Team:Groningen/protocols/biofilm media

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The strains were grown overnight at 37°C on LB broth.
Next, 1ml of liquid culture was used to inoculate every 10ml MSgg medium. this in turn was incubated at 27 C for 5 days.

Media for macrocolony formation


2x SG medium (100mL)
compound amount treatment
Nutrient broth (Difco) 1.6g
KCl 0.2g
MgSO47H2O 0.05g
agar 1.5g
add al together and autoclave
Ca(NO3)24H2O 1mM 0.1mL
MnCl24H2O 0.1mM 0.1mL
FeSO4 1 uM 0.1mL
0.1% glucose 0.5mL

LB/Mn/glucose medium (100mL)
compound amount treatment
LB/ 1.5% agar 100ml autoclave
MnCl24H2O 0.1mM 0.1mL
0.1% glucose 0.5mL

MSgg medium (100mL)
compound amount treatment
KH2PO4 0.026g
K2HPO4 0.061g
MOPS 100mM 2.09g
MgCl26H2O 2mM 0.04g
agar 1.5g
add al together and autoclave
CaCl2 700mM 0.1mL
MnCl2 50uM 50uL
FeCl3 50 uM 0.1mL
ZnCl2 1 uM 0.1mL
thiamine 2 uM 0.1mL
0.5% glycerol 0.57mL
0.5% glutamate 10mL
tryptophan 1mL


Grow cells overnight in LB / glass tubes at 37C.
Inoculate 2uL spot on plate dry 15 minutes at 30C
Inoculate 30C 2-4 days