Compound | Amount | Unit | |||
---|---|---|---|---|---|
|
Beef extract | 5.0 | g | ||
|
Trypticase
(BBL) |
5.0 | g | ||
|
Yeast extract | 10.0 | g | ||
|
Mineral solution | 40.0 | ml | ||
|
(NH4)2SO4 | 0.5 | g | ||
|
Glucose | 5.0 | g | ||
|
Resazurin | 1.0 | mg | ||
|
Haemin
(dissolved in 0.5 ml 1 N NaOH) |
5.0 | mg | ||
|
Vitamin K1
(0.5% sol. in ethanol) |
0.2 | ml | ||
|
L-Cysteine HCl x H2O | 0.5 | g | ||
|
Distilled water | 960.0 | ml | ||
|
Bovine serum
(made anaerobic by preincubation in a GasPak container over-night) |
5.0 | % | ||
1 Dissolve ingredients except cysteine-HCl in water, boil for 5 min, cool to room temperature while gassing with 100% CO2, add cysteine-HCl and adjust pH to 6.9 with 8 N NaOH while gassing is continued. Switch to 100% N2 gas after pH has reached the desired value, distribute in anaerobic tubes under nitrogen, and autoclave. | |||||
2 Adjust pH to 6.9. | |||||
3 Before use add 5% (v/v) bovine serum (made anaerobic by preincubation in a GasPak container over-night). |
Compound | Amount | Unit | |
---|---|---|---|
|
CaCl2 x 2 H2O | 0.30 | g |
|
MgSO4 x 7 H2O | 0.48 | g |
|
K2HPO4 | 1.00 | g |
|
KH2PO4 | 1.00 | g |
|
NaHCO3 | 10.00 | g |
|
NaCl | 2.00 | g |
|
Distilled water | 1000.00 | ml |