Compound | Amount | Unit | |||
---|---|---|---|---|---|
|
HEPES
(SIGMA) |
11.92 | g | ||
|
KH2PO4 | 0.30 | g | ||
|
NaCl | 0.60 | g | ||
|
MgCl2 x 6 H2O | 0.10 | g | ||
|
CaCl2 x 2 H2O | 0.08 | g | ||
|
Trace element solution | 10.00 | ml | ||
|
NH4Cl | 1.00 | g | ||
|
Sodium resazurin
(0.1% w/v) |
0.50 | ml | ||
|
KHCO3 | 2.00 | g | ||
|
2-Furoic acid
(SIGMA) |
2.24 | g | ||
|
Wolin's vitamin solution (10x) | 1.00 | ml | ||
|
L-Cysteine HCl x H2O | 0.30 | g | ||
|
Na2S x 9 H2O | 0.30 | g | ||
|
Distilled water | 1000.00 | ml | ||
1 Dissolve ingredients except bicarbonate, vitamins, furoic acid, cysteine and sulfide and adjust pH of solution to 7.0. Sparge medium with 80% N2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Dispense medium under the same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Add furoic acid (stock solution neutralized with NaOH), cysteine and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas and bicarbonate from a sterile anoxic stock solution prepared under 80% N2 and 20% CO2 gas mixture. Prior to use adjust pH of complete medium to 7.0 - 7.2, if necessary. |
Compound | Amount | Unit | |||
---|---|---|---|---|---|
|
Nitrilotriacetic acid
(NTA) |
12.80 | g | ||
|
FeCl2 x 4 H2O | 1.00 | g | ||
|
MnCl2 x 4 H2O | 0.10 | g | ||
|
CoCl2 x 6 H2O | 0.03 | g | ||
|
CaCl2 x 2 H2O | 0.10 | g | ||
|
ZnCl2 | 0.10 | g | ||
|
CuCl2 | 0.02 | g | ||
|
H3BO3 | 0.01 | g | ||
|
Na2MoO4 x 2 H2O | 0.03 | g | ||
|
NiCl2 x 6 H2O | 0.10 | g | ||
|
NaCl | 1.00 | g | ||
|
Na2SeO3 x 5 H2O | 0.03 | g | ||
|
Na2WO4 x 2 H2O | 0.04 | g | ||
|
Distilled water | 1000.00 | ml | ||
1 First dissolve NTA in 200 ml of distilled water and adjust pH to 6.5 with KOH, then dissolve mineral salts. Finally adjust pH to 6.5 with KOH and make up to 1000.00 ml. |
Compound | Amount | Unit | |
---|---|---|---|
|
Biotin | 20 | mg |
|
Folic acid | 20 | mg |
|
Pyridoxine hydrochloride | 100 | mg |
|
Thiamine HCl | 50 | mg |
|
Riboflavin | 50 | mg |
|
Nicotinic acid | 50 | mg |
|
Calcium D-(+)-pantothenate | 50 | mg |
|
Vitamin B12 | 1 | mg |
|
p-Aminobenzoic acid | 50 | mg |
|
(DL)-alpha-Lipoic acid | 50 | mg |
|
Distilled water | 1000 | ml |