1027 .
METHANOGENS SALINE WATER MEDIUM
What is this page?
Equipment needed:
Anaerobic workbench
Autoclave
Filter
Hungate tubes
Final pH:
n.d.
Select a strain (optional):
ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
NH4Cl | 0.54 | g | ||
|
|
KH2PO4 | 0.14 | g | ||
|
|
MgCl2 x 6 H2O | 3.50 | g | ||
|
|
K2HPO4 | 0.30 | g | ||
|
|
CaCl2 x 2 H2O | 0.15 | g | ||
|
|
NaCl | 5.00 | g | ||
|
|
NaHCO3 | 2.52 | g | ||
|
|
Trace mineral solution | 1.00 | ml | ||
|
|
Se/W solution | 1.00 | ml | ||
|
|
Trace vitamins solution | 1.00 | ml | ||
|
|
Resazurin | 1.00 | mg | ||
|
|
Distilled water | 1000.00 | ml | ||
| 1 Mix components thoroughly and distribute the medium into culture vessels (e.g., 20 ml medium in 100 ml serum bottles or 5.0 ml medium in Hungate tubes) under a stream of H2-CO2 (80:20, v/v), seal with butyl rubber stoppers and autoclave. Allow to stand the autoclaved medium overnight. Add per liter the following solution from a sterile anaerobic stock (filter-sterilized and stored under a N2 atmosphere): | |||||
|
|
Trimethylamine
(1 M) |
10.00 | ml | ||
| 2 Prior to inoculation, add per liter the following solutions aseptically and anaerobically (autoclaved and stored under a N2 atmosphere): | |||||
|
|
L-Cysteine HCl x H2O
(5%) |
10.00 | ml | ||
|
|
Na2S x 9 H2O
(5%) |
10.00 | ml | ||
| 3 Comments: For strain JCM 19935, replace trimethylamine with yeast extract (final 0.1 g/L) and pressurize to 100 kPa H2-CO2 (80:20, v/v). For strain JCM 19936, prepare the medium without trimethylamine. Pressurize inoculated culture vessels to 100 kPa H2-CO2 (80:20, v/v). | |||||
Trace mineral solution #
1000
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
FeCl2 x 4 H2O | 1.990 | g |
|
|
MnCl2 x 4 H2O | 0.198 | g |
|
|
CoCl2 x 6 H2O | 0.130 | g |
|
|
ZnCl2 | 0.136 | g |
|
|
CuCl2 x 2 H2O | 1.700 | mg |
|
|
H3BO3 | 6.200 | mg |
|
|
Na2MoO4 x 2 H2O | 0.024 | g |
|
|
NiCl2 x 6 H2O | 0.013 | g |
|
|
AlCl3 | 0.013 | g |
|
|
Distilled water | 1000.000 | ml |
Se/W solution #
1000
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
Na2SeO3 x 5 H2O | 1.7 | mg |
|
|
Na2WO4 x 2 H2O | 3.3 | mg |
|
|
Distilled water | 1000.0 | ml |
Trace vitamins solution #
1000
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
Biotin | 4.9 | mg |
|
|
Folic acid | 8.8 | mg |
|
|
Pyridoxine hydrochloride | 4.1 | mg |
|
|
Thiamine HCl | 6.7 | mg |
|
|
Riboflavin | 7.5 | mg |
|
|
Nicotinic acid | 2.5 | mg |
|
|
DL-Calcium pantothenate | 9.5 | mg |
|
|
Vitamin B12 | 27.1 | mg |
|
|
p-Aminobenzoic acid | 2.7 | mg |
|
|
Lipoic acid | 4.1 | mg |
|
|
Distilled water | 1000.0 | ml |