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Stocks |
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5X Beijerinck's Salt Stock: |
|
| NH4Cl |
40 g |
| CaCl2 (2H2O) |
5 g |
| MgSO4 (2H2O) |
10 g |
To 1 L Distilled H20
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Phosphate Stock: |
|
| K2HPO4 (anhydrous) |
14.34 g |
| KH2PO4 (anhydrous) |
7.26 g |
| To 1 L Distilled H20 |
| Check pH shall be around pH to 7.1, adjust with KOH |
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TRIS Stock: |
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| Tris base |
242 g |
| to 1 L Distilled H20 |
| Titrate to pH 6.6 with HCl |
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Trace Elements Solution (Hutner 1946, J. Bact. 52: 213): |
|
| Na2EDTA |
50.0 g |
| H3BO3 (Boric acid) |
11.14 g |
| ZnSO4-7H2O |
22.0 g |
| MnCl2-4H2O |
5.1 g |
| FeSO4-7H2O |
5.0 g |
| CoCl2-6H2O |
1.6 g |
| CuSO4-5H2O |
1.6 g |
| (NH4)6Mo7O24-4H2O |
1.1 g |
| To 1 L Distilled H20 |
Follow the instructions:
- Add elements one at a time (except EDTA) to 550 mL milliQ H2O in a 1L Erlenmeyer flask.
- Heat the solution to approximately 70 °C.
- In a different beaker, add EDTA to 250 mL milliQ H2O and heat until dissolved.
- Add EDTA solution to salt solution (not vice versa) and bring the combined solution to a boil.
- Let the solution cool down and maintain temperature at 70-75 °C.
- Adjust pH to 6.5-6.8 with 20% KOH. Be careful: do not let the temperature fall below 70 °C or let the pH go over 6.8 otherwise you have to re-start.
- Dilute to 1000 mL.
- Use a cotton plug to cover flask (not parafilm!) and let it stand for two weeks until the colour changes from green to purple. Stir solution to help it, all along.
- Filter out red-brown precipitate and store in refrigerator. Note if there is a precipitate, then you don't know the real concentrations of elements in your solution.
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Agar Plates |
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Add 1.5-2% w/v of Noble Agar to medium (typical 6-8 g to 400 mL TAP solution)
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TAP receipt in PDF format: Tap.pdf |
