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