Medium data

Search for medium no. = [221].
Unless otherwise stated, sterilize media by autoclaving at 121°C for 15 min.

221   THERMOANAEROBACTER ETHANOLICUS MEDIUM

KH2PO4  1.5     g
Na2HPO4·12H2O  4.2     g
NH4Cl  0.5     g
MgCl2·6H2O  0.18    g
Yeast extract (BD-Difco)  2.0     g
Glucose  8.0     g
Vitamin solution (see below)  0.5     ml
Wolfe's modified mineral elixir (see below)  5.0     ml
Resazurin  1.0     mg
Reducing solution (see below)  40.0     ml
Distilled water  960.0     ml
Mix ingredients, except reducing solution, and bring to the boil while gassing with N2-H2 (95:5) gas mixture. When the color turns from blue to reddish pink, add the reducing solution during continuous gassing and boiling. Anaerobically dispense the medium in tubes under a N2-H2 (95:5) atmosphere. Stopper the tubes and autoclave.
Vitamin solution:
Biotin  20.0     mg
p-Aminobenzoic acid  50.0     mg
Folic acid  20.0     mg
Calcium pantothenate  50.0     mg
Nicotinic acid  50.0     mg
Vitamin B12  1.0     mg
Thiamine·HCl  5.0     mg
Pyridoxine·HCl  100.0     mg
Thioctic acid  50.0     mg
Riboflavin  5.0     mg
Distilled water  500.0     ml
Store at -20C or 4C in the absence of light.
Wolfe's modified mineral elixir:
Nitrilotriacetic acid  1.5     g
MgSO4·7H2O  3.0     g
MnSO4·xH2O  0.5     g
NaCl  1.0     g
FeSO4·7H2O  0.1     g
Co(NO3)2·6H2O  0.1     g
CaCl2  0.1     g
ZnSO4·7H2O  0.1     g
CuSO4·5H2O  0.01    g
AlK(SO4)2  0.01    g
H3BO3  0.01    g
Na2MoO4·2H2O  0.01    g
Na2SeO3  1.0     mg
Distilled water  1.0     L
Dissolve nitrilotriacetic acid and adjust pH to 6.5 with KOH. Then proceed to add minerals in the order listed.
Reducing solution:
0.2 N NaOH  200.0     ml
Na2S·9H2O  2.5     g
L-Cysteine·HCl·H2O  2.5     g
Boil NaOH solution and cool down under a N2-H2 (95:5) atmosphere, then add Na2S·9H2O and L-cysteine·HCl·H2O to the solution. Autoclave it under a N2-H2 (95:5) atmosphere.