ATCC medium: 1768 Geobacter metallireducens Medium
Ferric citrate (Sigma F-6129).......................13.7 g
Wolfe's Vitamin Solution (see below)..........10.0 ml
Wolfe's Mineral Solution (see below)..........10.0 ml
NaHCO3 .......................................................2.5 g
NH4Cl ...........................................................0.25 g
NaH2PO4 . H2O.............................................0.6 g
KCl................................................................0.1 g
Sodium acetate.............................................6.8 g
Distilled, deionized water……......................1.0 L
Heat about 400 ml of water on a hot stir plate to near boiling. Add ferric citrate, allow it to dissolve, then cool the solution to room temperature in a slurry of ice. Add 400 ml of water; this quickly cools the medium to room temperature. Adjust the pH to 6.0 using 10 N NaOH; when the pH approaches 5.0, add NaOH by drops until the pH level is established. Approximately 6 ml of 10 N NaOH per liter will be needed per liter of medium. Add the remaining ingredients and bring the final volume up to 1.0 L with water. Bubble the medium with 80% N2 and 20% CO2. The final pH should be 6.8-7.0.‘
Do not expose this medium to sunlight.
Wolfe's Vitamin Solution: Available from ATCC as a sterile ready-to-use liquid (Vitamin Supplement, catalog no. MD-VS).
Biotin..............................................2.0 mg
Folic acid........................................2.0 mg
Pyridoxine hydrochloride..............10.0 mg
Thiamine . HCl...............................5.0 mg
Riboflavin.......................................5.0 mg
Nicotinic acid.................................5.0 mg
Calcium D-(+)-pantothenate.........5.0 mg
Vitamin B12....................................0.1 mg
p-Aminobenzoic acid....................5.0 mg
Thioctic acid..................................5.0 mg
Distilled water................................1.0 L
Wolfe's Mineral Solution: Available from ATCC as a sterile ready-to-use liquid (Trace Mineral Supplement, catalog no. MD-TMS.) Nitrilotriacetic acid................1.5 g
MgSO4 . 7H2O .......................3.0 g
MnSO4 . H2O ........................0.5 g
NaCl......................................1.0 g
FeSO4 . 7H2O .......................0.1 g
CoCl2 . 6H2O ........................0.1 g
CaCl2 ....................................0.1 g
ZnSO4 . 7H2O .......................0.1 g
CuSO4 . 5H2O .......................0.01 g
AlK(SO4)2 . 12H2O.................0.01 g
H3BO3 ....................................0.01 g
Na2MoO4 . 2H2O....................0.01 g
Distilled water........................1.0 L
Add nitrilotriacetic acid to approximately 500 ml of water and adjust to pH 6.5 with KOH to dissolve the compound. Bring volume to 1.0 L with remaining water and add remaining compounds one at a time.