Protocols
Chlamydomonas Protocol
Cultivation
Media
To be able to work with C. reinhardtii, you will need medium. This medium can be in either liquid or solid form and is essential for any experiment regarding C. reinhardtii. The media we used for C. reinhardtii is TAP medium.
Media | 0,5 L | 1,8L |
---|---|---|
40 x TAP/TA/TAPi | 12,5 ml | 20 ml |
40 x Beijerink Salts | 12,5 ml | 20 ml |
Trace Elements (revised) | 3,5 ml | 5,6 ml |
H2O | 471,5 ml | 754,4 ml |
For Plates: + Agar (1,8 %) | 7,2 g | 14,4 g |
Tap Medium should be adjusted to 7,0 - 7,2 pH and autoclaved.
Stocks
40 x TAP 1L | autoclave, store at 4°C |
---|---|
Tris (121,14 g/mol) | 96,8 g (0,8 M) |
1 M (K) PO4 | 40 ml (40 mM) |
adjust pH 7.0-7.2 with conc. AcOH (~44ml)
40 x TA 1L | autoclave, store at 4°C |
---|---|
Tris (121,14 g/mol) | 96,8 g (0,8 M) |
adjust pH 7.0-7.2 with conc. AcOH (~44ml)
40 x TAPi 1L | autoclave, store at 4°C |
---|---|
Tris (121.14 g/mol) | 96,8 g (0,8 M) |
1 M K2HPO3 120.08 g/mol | 40ml (40mM) |
adjust pH 7.0-7.2 with conc. AcOH (~44ml)
1 M (K)PO4 | MW(g/mol) | conc. | 420 ml |
---|---|---|---|
K2HPO4 ∙3 H2O | 228,22 | 1 M; 228 g/l | ≈0,6 M; 136 g/l |
K2HPO4 | 136,09 | 1 M; 136 g/l | ≈ 0,4 M; 55,08 g/l |
250 ml 1 M K2HPO4 + ca. 170 ml KH2PO4 , adjust to pH 7, autoclave, store at 4°C
1 M (K)PO3 | MW(g/mol) | conc. | 1 l |
---|---|---|---|
K2HPO3 | 120.086 | 1 M; 120.086 g/l | ≈1 M; 120.086 g/l |
K2HPO3 was obtained by Haihang co. ltd. (13977-65-6) Autoclave, store at 4°C
40 x Beijerinck salts 1 L | autoclave, store at 4°C |
---|---|
NH 4Cl | 16 g |
CaCl2 ∙2H 2O | 2 g |
MgSO 4 ∙7H 2O | 4 g |
Trace Elements (revised)
To make the trace elements following publication was used: Kropat, J., Hong-Hermesdorf, A., Casero, D., Ent, P., Castruita, M., Pellegrini, M., … Malasarn, D. (2011). A revised mineral nutrient supplement increases biomass and growth rate in Chlamydomonas reinhardtii. The Plant Journal, 66(5), 770–780. http://doi.org/10.1111/j.1365-313X.2011.04537.x
Additives
Additive (optional) | Working Concentration |
---|---|
Arginin | 100 µg/ml |
Paromomycin | 10-12 µg/ml |
Hygromycin | 10 µg/ml |
Ampicillin | 100-500 µg/ml |