1287 .
NATRONOANAEROARCHAEA MEDIUM WITH YEAST EXTRACT AND GLUCOSE
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Equipment needed:
Anaerobic workbench
Autoclave
Filter
Final pH:
n.d.
Select a strain (optional):
ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Base salt medium | 750.0 | ml | ||
|
|
Base soda medium | 250.0 | ml | ||
| 1 Mix two media autoclaved separately. Aseptically add the following solutions (autoclaved or *filter-sterilized): | |||||
|
|
MgSO4
(1 M) |
1.0 | ml | ||
|
|
Trace element solution | 1.0 | ml | ||
|
|
Selenite-tungstate solution
(*) |
1.0 | ml | ||
|
|
Trace vitamins
(*) |
5.0 | ml | ||
| 2 To complete the medium, aseptically and anaerobically distribute the medium into culture vessels containing steam-sterilized sulfur (see below, approximately 5.0 g/L) under an argon gas stream, and seal with butyl rubber stoppers. Then, add per liter the following solutions (autoclaved or *filter-sterilized): | |||||
|
|
Yeast extract
(10%) |
1.0 | ml | ||
|
|
Glucose
(*, 1.0 M) |
10.0 | ml | ||
|
|
Na2S x 9 H2O
(5%) |
2.5 | ml | ||
Sterilization of sulfur #
| 1 Steam sulfur for 3 hr, or autoclave at 105C for 1 hr, on each of 3 successive days. |
Base salt medium #
Base soda medium #
Trace element solution #
1000
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
EDTA | 5.00 | g | ||
|
|
FeSO4 x 7 H2O | 2.00 | g | ||
|
|
ZnSO4 x 7 H2O | 0.10 | g | ||
|
|
MnCl2 x 4 H2O | 0.03 | g | ||
|
|
H3BO3 | 0.30 | g | ||
|
|
CoCl2 x 6 H2O | 0.20 | g | ||
|
|
CuCl2 x 2 H2O | 0.01 | g | ||
|
|
NiCl2 x 6 H2O | 0.02 | g | ||
|
|
Na2MoO4 x 2 H2O | 0.03 | g | ||
|
|
Distilled water | 1000.00 | ml | ||
| 1 Adjust pH to 3.6. | |||||
Selenite-tungstate solution #
1000
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
NaOH | 0.4 | g |
|
|
Na2SeO3 x 5 H2O | 6.0 | mg |
|
|
Na2WO4 x 2 H2O | 8.0 | mg |
|
|
Distilled water | 1000.0 | ml |
Trace vitamins #
1000
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
Biotin | 2.0 | mg |
|
|
Folic acid | 2.0 | mg |
|
|
Pyridoxine hydrochloride | 10.0 | mg |
|
|
Thiamine HCl | 5.0 | mg |
|
|
Riboflavin | 5.0 | mg |
|
|
Nicotinic acid | 5.0 | mg |
|
|
Calcium pantothenate | 5.0 | mg |
|
|
Vitamin B12 | 0.1 | mg |
|
|
p-Aminobenzoic acid | 5.0 | mg |
|
|
Lipoic acid | 5.0 | mg |
|
|
Distilled water | 1000.0 | ml |