ATCC medium: 2332 Thermotoga petrophila medium
Instant Ocean...........................26.0 g
(NH4)2SO4.................................1.0 g
KH2PO4 ...................................0.5 g
Yeast extract............................2.0 g
MOPS (Sigma).............................5.0 g
Wolfe's Mineral Solution (see below)....10.0 ml
Wolfe's Vitamin Solution (see below)....10.0 ml
Resazurin................................0.5 mg
Na2S . 9H2O ..............................0.3 g
Distilled water........................980.0 ml
Dissolve ingredients (except sodium sulfide and vitamin solution) boil medium for 3 minutes, then cool to room temperature under N2.
Dispense under same gas in anaerobic vessels and autoclave.
The final pH of the completed medium should be 7.0.
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.
2 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