Used for Chlamydomonas as well as other species.
Reference: Sueoka, N., Chiang, K.S. and Kates, J.R. (1967) Deoxyribonucleic acid replication in meiosis of Chlamydomonas reinhardtii. Isotopic transfer experiments with a strain producing eight zoospores. J. Mol. Biol. 25: 44-67.
| Stock | Substance | Concentration | Amount added to medium (mL/L) |
|---|---|---|---|
| 1. | Beijerinck's #2: |
50 mL |
|
| NH4Cl | 5 g/500 mL | ||
| MgSO4•7H2O | 0.2 g/500 mL | ||
| CaCl2•2H2O | 0.1 g/500 mL | ||
| 2. | Phosphate #2: |
25 mL |
|
| KH2PO4 | 14.8 g/500 mL | ||
| K2HPO4 | 28.8 g/500 mL | ||
| 3. | AAP trace metal solution | See below | 1 mL |
Final pH of medium is 6.8 and if necessary can be adjusted using 1N HCl and 1N NaOH.
Autoclave for 20 minutes or longer if using larger quantities than 1 Litre. Allow medium to stand overnight before use.
Trace metal solution
| Substance | Concentration (mg/500 mL) |
|---|---|
| 1. H3BO3 | 92.76 mg |
| 2. MnCl2•4H2O | 207.69 mg |
| 3. ZnCl2 | 1.64 mg |
| 4. FeCl3•6H2O | 79.89 mg |
| 5. Na2EDTA•2H2O | 150.00 mg |
| 6. CoCl2•6H2O | 1.30 mg |
| 7. Na2MoO4•2H2O | 3.63 mg |
| 8. CuCl2•2H2O | 0.006 mg |
Dissolve each ingredient before adding the next. Allow to stand until a precipitate develops. Filter into a sterile bottle when colour changes from greenish-brown to purple. Store in refrigerator.