Team:Groningen/protocols/biofilm media

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The strains were grown overnight at 37°C on LB broth.
Next, 1 ml of liquid culture was used to inoculate every 10 ml 2x Sgg medium. this in turn was incubated at 27°C for 5 days.
Sgg is 2x SG supplemented with 1% [wt/vol] glycerol.

Media for macrocolony formation


2x SG medium (100 mL)
compound amount treatment
Nutrient broth (Difco) 1.6 g
KCl 0.2 g
MgSO4.7H2O 0.05 g
agar 1.5 g
add everything together and autoclave
Ca(NO3)2.4H2O 1 mM 0.1 mL
MnCl2.4H2O 0.1 mM 0.1 mL
FeSO4 1 µM 0.1 mL
0.1% glucose 0.5 mL

LB/Mn/glucose medium (100 mL)
compound amount treatment
LB/ 1.5% agar 100 ml autoclave
MnCl2.4H2O 0.1 mM 0.1mL
0.1% glucose 0.5 mL

MSgg medium (100 mL)
compound amount treatment
KH2PO4 0.026 g
K2HPO4 0.061 g
MOPS 100 mM 2.09 g
MgCl2.6H2O 2 mM 0.04 g
agar 1.5 g
add everything together and autoclave
CaCl2 700 mM 0.1 mL
MnCl2 50 µM 50 µL
FeCl3 50 µM 0.1 mL
ZnCl2 1 µM 0.1 mL
thiamine 2 µM 0.1 mL
0.5% glycerol 0.57 mL
0.5% glutamate 10 mL
tryptophan 1 mL


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