1444 .
PIRELLULA MEDIUM (M30a)
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Equipment needed:
Autoclave
Filter
Final pH:
7.5
Select a strain (optional):
Solution 1 #
950
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Hutner's salts | 20 | ml | ||
|
|
Tris-HCl buffer
(0.1M, pH 7.5) |
50 | ml | ||
|
|
Artifical sea water | 500 | ml | ||
|
|
Agar | 18 | g | ||
|
|
Distilled water | 380 | ml | ||
| 1 Adjust pH to 7.5, autoclave at 121°C for 20 min. | |||||
| 2 For agar plates increase the pH of the Tris/HCl buffer to 8.5. | |||||
Solution 2 #
50
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
N-Acetylglucosamine | 2.0 | g | ||
|
|
Na2HPO4 x 2 H2O | 0.1 | g | ||
|
|
Vitamin solution (double conc.) | 10.0 | ml | ||
|
|
Distilled water | 40.0 | ml | ||
| 1 Adjust pH to 7.5, filter-sterilize, and add to solution 1. | |||||
Hutner's salts #
1000
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Nitrilotriacetic acid
(NTA) |
10.000 | g | ||
|
|
MgSO4 x 7 H2O | 29.700 | g | ||
|
|
CaCl2 x 2 H2O | 3.335 | g | ||
|
|
(NH4)6Mo7O24 x 4 H2O | 9.250 | mg | ||
|
|
FeSO4 x 7 H2O | 99.000 | mg | ||
|
|
"Metals 44" | 50.000 | ml | ||
|
|
Distilled water | 950.000 | ml | ||
| 1 Dissolve the nitrilotriacetic acid. | |||||
| 2 Adjust pH to 7.0 with KOH (~7.3 g). | |||||
| 3 Dissolve other salts separately and combine. | |||||
| 4 Adjust pH to 6.8 with NaOH or H2SO4. | |||||
Artifical sea water #
1000
Vitamin solution (double conc.) #
1000
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Biotin | 4.0 | mg | ||
|
|
Folic acid | 4.0 | mg | ||
|
|
Pyridoxine hydrochloride | 20.0 | mg | ||
|
|
Riboflavine | 10.0 | mg | ||
|
|
Thiamine-HCl x 2 H2O | 10.0 | mg | ||
|
|
Nicotinamide | 10.0 | mg | ||
|
|
D-Calcium pantothenate | 10.0 | mg | ||
|
|
Vitamin B12 | 0.2 | mg | ||
|
|
p-Aminobenzoic acid | 10.0 | mg | ||
|
|
Distilled water | 1000.0 | ml | ||
| 1 Store in the dark and cold (5°C). | |||||
"Metals 44" #
1000
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Na-EDTA | 250.0 | mg | ||
|
|
ZnSO4 x 7 H2O | 1095.0 | mg | ||
|
|
FeSO4 x 7 H2O | 500.0 | mg | ||
|
|
MnSO4 x H2O | 154.0 | mg | ||
|
|
CuSO4 x 5 H2O | 39.2 | mg | ||
|
|
Co(NO3)2 x 6 H2O | 24.8 | mg | ||
|
|
Na2B4O7 x 10 H2O | 17.7 | mg | ||
|
|
Distilled water | 1000.0 | ml | ||
| 1 Dissolve the EDTA and add a few drops of concentrated H2SO4 to retard precipitation of the heavy metal ions. | |||||