Protocols
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Chlamydomonas Protocol
Media & Plates
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
Cultivation
Transformation
TODO
Colony Screening PCR
TODO
E. coli Protocols
Cultivation
Media & Plates
TODO
Overnight Cultures
TODO
Transformation
TODO
DNA-Isolation
TODO
Synthesis Protocols
Sequence & Primer Design
TODO
PCR
TODO
Digestion
TODO
Ligation
TODO
General
Gel Electrophoresis
TODO
Protein Expression: SDS-page
TODO
Workflow
Plasmid assembly
TODO