ATCC medium: 1465 Basal thermophile medium
Solution 1:
NH4Cl ....................................0.9 g
MgCl2 . 6H2O .............................0.2 g
KH2PO4 ...................................0.75 g
K2HPO4 ...................................1.5 g
Trace Element Solution (see below).......9.0 ml
10% FeSO4 . 7H2O .........................0.03 ml
0.2% Resazurin...........................1.0 ml
Wolfe's Vitamin Solution (see below).....5.0 ml
Trypticase Peptone (BD 211921)..........10.0 g
Distilled water........................850.0 ml
Solution 2:
Yeast extract............................3.0 g
Distilled water........................100.0 ml
Solution 3:
Glucose..................................5.0 g
Distilled water.........................50.0 ml
Autoclave the above solutions separately at 121C for 45 minutes.
Combine solutions under 100% N2 and dispense aseptically using the same gas phase.
At time of inoculation add 10% sterile Na2S . 9H2O to a final concentration of 0.1%.
Trace Element Solution:
Nitrilotriacetic acid (Neutralize to pH 6.5 with KOH).....12.5 g
FeCl3 . 4H2O ...............................................0.2 g
MnCl2 . 4H2O ...............................................0.1 g
CoCl2 . 6H2O ...............................................0.017 g
CaCl2 . 2H2O ...............................................0.1 g
ZnCl2 ......................................................0.1 g
CuCl2 ......................................................0.02 g
H3BO3 ......................................................0.01 g
Na2MoO4 . 2H2O..............................................0.01 g
NaCl.......................................................1.0 g
Na2SeO3 ....................................................0.02 g
Distilled water............................................1.0 L
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