Compound | Amount | Unit | |||
---|---|---|---|---|---|
|
Solution A | 960 | ml | ||
|
Solution B | 20 | ml | ||
|
Solution C | 20 | ml | ||
1 Dissolve ingredients of solution A (except cysteine) and sparge medium with 100% N2 gas for 30 - 45 min to make it anoxic. Add cysteine and adjust pH 7.2 with KOH, then distribute under the same gas atmosphere in anoxic Hungate-type tubes or serum vials and autoclave. Solution B is autoclaved separately under 80% N2 and 20% CO2 gas atmosphere and solution C is autoclaved under 100% N2 gas atmosphere. To complete the medium appropriate amounts of solutions B and C are added to the sterile solution A in the sequence as indicated. Final pH of the complete medium should be 7.5. | |||||
2 For solid medium add 10.00 g/l agar (BD Bacto) to solution A. |
Compound | Amount | Unit | |
---|---|---|---|
|
CaCl2 x 2 H2O | 0.04 | g |
|
MgSO4 x 7 H2O | 0.50 | g |
|
Yeast extract
(OXOID) |
4.00 | g |
|
D-Glucose | 2.00 | g |
|
Sodium resazurin
(0.1% w/v) |
0.50 | ml |
|
L-Cysteine HCl x H2O | 0.50 | g |
|
Agar
, for solid medium
optional |
10.00 | g |
|
Distilled water | 960.00 | ml |