1095 .
BICARBONATE BUFFERED MEDIUM
What is this page?
Equipment needed:
Anaerobic workbench
Autoclave
Filter
Final pH:
n.d.
Select a strain (optional):
ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Na2HPO4 x 2 H2O | 0.53 | g | ||
|
|
KH2PO4 | 0.41 | g | ||
|
|
NH4Cl | 0.30 | g | ||
|
|
NaCl | 0.30 | g | ||
|
|
FeCl2 solution | 1.00 | ml | ||
|
|
Trace element solution | 1.00 | ml | ||
|
|
Selenite-tungstate solution | 0.50 | ml | ||
|
|
Yeast extract | 0.10 | g | ||
|
|
Resazurin | 0.50 | mg | ||
|
|
Distilled water | 930.00 | ml | ||
| 1 Mix components and autoclave under a N2-CO2 (4:1, v/v) gas mixture. After cooling, add the following solutions (autoclaved or *filter-sterilized): | |||||
|
|
MgCl2 x 6 H2O
(10%) |
1.00 | ml | ||
|
|
CaCl2 x 2 H2O
(10%) |
1.00 | ml | ||
|
|
Trace vitamins
(*) |
10.00 | ml | ||
|
|
NaHCO3
(*, 8%) |
50.00 | ml | ||
|
|
Substrate Solution | 10.00 | ml | ||
| 2 Aseptically and anaerobically distribute the medium into culture vessels (e.g., 50 ml of the medium in 120 ml serum bottle) under a N2-CO2 (4:1, v/v) gas mixture, and seal with butyl rubber stoppers. Prior to use, add per liter the following solution (autoclaved and stored under a N2 atmosphere): | |||||
|
|
Na2S x 9 H2O
(5%) |
10.00 | ml | ||
Substrate Solution #
| 1 For strain JCM 30818, 1 M glycerin solution; | |||||
| 2 For strain JCM 31247, 1 M trisodium citrate solution; | |||||
| 3 For strain JCM 31248, 1 M maltose solution. |
FeCl2 solution #
1000
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
HCl
(7.7 M, 25%) |
10.0 | ml |
|
|
FeCl2 x 4 H2O | 1.5 | g |
|
|
Distilled water | 990.0 | ml |
Trace element solution #
1000
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
ZnCl2 | 70 | mg |
|
|
MnCl2 x 4 H2O | 100 | mg |
|
|
H3BO3 | 6 | mg |
|
|
CoCl2 x 6 H2O | 190 | mg |
|
|
CuCl2 x 2 H2O | 2 | mg |
|
|
NiCl2 x 6 H2O | 24 | mg |
|
|
Na2MoO4 x 2 H2O | 36 | mg |
|
|
Distilled water | 1000 | ml |
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 |