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Revision as of 18:20, 1 November 2017
Experiments
Chlamydomonas reinhardtii growth
The strain that will be utilized is cell wall deficient, the used in [1] is the strain cw15-30-derived UVM4 C. The culture medium in which they grow are TAP (solid) and TAPS (liquid). Light stimulation will be required, for which a structure capable of covering the Erlenmeyer flasks or the petri dishes in which C. reinhardtii would be growing while uniformly illuminating with the whole visible light spectrum will be designed and built. In [1] the lamp that is put into use has the following characteristics: Nano Light, 11 Watts, Dennerle, Vinningen, Germany, which provided constant light with the following details: 2500 lux, eq. 72.5 μE/m2∙s1.
For the construction of calibration curves the medium to be used is TAPS without acetate.
[1] Chávez, M. N., Schenck, T. L., Hopfner, U., Centeno-Cerdas, C., Somlai-Schweiger, I., Schwarz, C., ... & Nickelsen, J. (2016). Towards autotrophic tissue engineering: photosynthetic gene therapy for regeneration. Biomaterials, 75, 25-36.
TAP (Tris-Acetate-Phosphate)
Solution/Compound |
Volume [ml] |
1M Tris base (e.g. Trizma) |
20 |
Phosphate Buffer II* |
1 |
Solution A |
10 |
Hutner's trace elements* |
1 |
Glacial Acetic Acid |
1 |
|
Compound | Mass [gr] |
---|---|
$$K_2 HPO_{4}$$ | 10.8 |
$$KH_2 PO_4$$ | 5.6 |
Compound | Mass [gr] |
---|---|
$$KH_4 Cl$$ | 20 |
$$MgSO_4\cdot7H_2O$$ | 5 |
$$CaCl_2\cdot2H_2O$$ | 2.5 |
Regarding Hutner´s trace elements:
Source: http://www.chlamycollection.org/methods/media-recipes/hutners-trace-elements/
TAPS
It’s TAP medium supplemented with 1% w/v sorbitol.
Sequences
poto