Reference: Jüttner et al., 1983.
This medium is used successfully for some cyanobacteria. Vitamins may be added. The CPCC currently adds f/2 vitamins to the Jüttner medium for all strains.
Stock | Substance | Concentration | Amount added to medium (mL/L) |
---|---|---|---|
1. | CaCl2•2H2O | 8.8 g/L | 10 mL |
NaNO3 | 68 g/L | ||
2. | K2HPO4* | 7.0 g/L* | 10 mL |
3. | MgSO4•7H2O | 9.9 g/L | 10 mL |
4. | NaFeEDTA | 367 mg/L | 10 mL |
5. | Trace metal solution | See below | 10 mL |
6. | f/2 vitamin solution | See below | 1 mL |
*May also prepare a 9.1 g/L stock solution of K2HPO4•3H2O instead of 7.0 g/L of K2HPO4
Adjust the pH of the medium to approximately 9 prior to autoclaving. However, please note that the CPCC has been pH adjusting the medium to approximately 8 (i.e., 7.5-8.5). Store stock solutions 1-5 in the fridge in the dark. It is especially important to keep Stock 4 in the dark, so use amber bottle and/or cover the bottle in tin foil. The f/2 vitamins (Stock 6) should be stored in the freezer in 1 mL aliquots and passively thawed as needed.
Trace metal solution
Add 100 mL Trace Elements Solution I plus 100 mL Trace Elements Solution II to 800 mL purified H2O.
Trace elements solution I:
Substance | Concentration (mg/L) |
---|---|
1. H3BO3 | 618 mg |
2. MnCl2•4H2O | 1979 mg |
3. ZnSO4•7H2O | 58 mg |
4. CuSO4•5H2O | 50 mg |
5. CoSO4•7H2O | 56 mg |
Trace elements solution II:
Substance | Concentration (mg/L) |
---|---|
1. Na2MoO4•2H2O | 484 mg |
Add each substance in the order that they appear here and ensure that each is visually dissolved prior to adding the next on the list. After all substances have been added the stock solution should be topped up with purified water to 1000 mL. Then combine 100 mL of Trace Elements Solution I with 100 mL of Trace Elements Solution II and top up to 1000 mL.
f/2 vitamin solution
Substock solutions:
First prepare separate substock solutions of cyanocobalamin (Vitamin B12) and biotin in purified water (e.g., Milli-Q, Nanopure, distilled water).
Substock | Substance | Concentration |
---|---|---|
1. | Vitamin B12 (Cyanocobalamin) | 10 mg/10 mL |
2. | Biotin | 5 mg/50 mL |
Working stock solution:
Then prepare the final working solution by combining the following amounts of the two substock solutions plus thiamine HCl to 100 mL of purified water.
Substance | Concentration (per 100 mL) |
---|---|
1. Vitamin B12 (Cyanocobalamin) | 0.1 mL |
2. Biotin | 1.0 mL |
3. Thiamine HCl | 20 mg |
Dispense the working solution as 1 mL aliquots in cryovials for convenient use when preparing the medium. Store the remainder of the primary stocks and working solution in polyethylene bottles. Wrap vials/bottles with Parafilm to avoid moisture loss and store all solutions frozen.
References: Guillard and Ryther 1962, Guillard 1975