843 .
LIND BHT MEDIUM
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Equipment needed:
Anaerobic workbench
Autoclave
Filter
Final pH:
n.d.
Select a strain (optional):
ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
KH2PO4 | 0.30 | g | ||
|
|
K2HPO4 | 0.30 | g | ||
|
|
NH4Cl | 1.00 | g | ||
|
|
NaCl | 20.00 | g | ||
|
|
KCl | 0.10 | g | ||
|
|
CaCl2 x 2 H2O | 0.10 | g | ||
|
|
Trace element solution | 1.00 | ml | ||
|
|
Yeast extract | 2.00 | g | ||
|
|
Na2S2O3 x 5 H2O | 4.96 | g | ||
|
|
L-Cysteine HCl x H2O | 0.50 | g | ||
|
|
Resazurin | 1.00 | mg | ||
|
|
Distilled water | 925.00 | ml | ||
| 1 Mix components thoroughly and autoclave under a N2-CO2 (4:1, v/v) gas mixture. After cooling, add the following solutions from anaerobic stocks (autoclaved or *filter-sterilized): | |||||
|
|
NaHCO3
(*, 8%) |
25.00 | ml | ||
|
|
MgCl2 x 6 H2O
(10%) |
30.00 | ml | ||
|
|
Sodium acetate
(1.0 M) |
20.00 | ml | ||
| 2 Then, aseptically distribute the medium into culture vessels under a stream of H2-CO2 (4:1, v/v) and seal with butyl rubber stoppers. Prior to inoculation, add per liter the following solution from an anaerobic stock (autoclave and store under a N2 atmosphere): | |||||
|
|
Na2S x 9 H2O
(5%) |
8.00 | ml | ||
| 3 Pressurize inoculated culture vessels to 200 kPa H2-CO2 (4:1, v/v). | |||||
Trace element solution #
1000
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
HCl
(25%) |
12.5 | ml |
|
|
FeSO4 x 7 H2O | 2.1 | g |
|
|
H3BO3 | 30.0 | mg |
|
|
MnCl2 x 4 H2O | 100.0 | mg |
|
|
CoCl2 x 6 H2O | 190.0 | mg |
|
|
NiCl2 x 6 H2O | 24.0 | mg |
|
|
CuCl2 x 2 H2O | 2.0 | mg |
|
|
ZnSO4 x 7 H2O | 144.0 | mg |
|
|
Na2MoO4 x 2 H2O | 36.0 | mg |
|
|
Distilled water | 987.0 | ml |