IGEM:UNAM/2009/Notebook/IGEM project/2011/05/05

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Medium of Chlamydomonas reinhardtii

Abstract

Today Miriam gave me the protocol to make TAP medium.

Tap Medium

Reagent Quantity
Beijerincks 20X 50 ml
Acetic acid 1 ml
Tris 2.42g
Phosphate solution 1M pH 7 1 ml
Trace Element 1ml

Gauging to 1 liter.


Beijerincks 20X solution

NH4Cl 8 g
CaCl2·H2O 1 g
MgSO4· 7H2O 2 g

Trace Element Solution

Reagent Quantity
EDTA sodium salt 50 g
ZnSO4·7H2O 22 g
H3BO3(a) 11.4 g
FeSO4·7H2O 4.99 g
MnCl2·H2O 5.06 g
CoCl·6H2O(b) 1.61 g
CuSO4·5H2O 1.57 g
(NH4)6Mo7O24·H2O 1. 10g

(a) Heat to dissolve (b) Shake to disolve

Dissolve each element independently.

- Boric acid in 200 ml. - Sulphate if zinc in 200 ml. - EDTA in 250 ml. - Other elements in 50 ml.

  • 1. Put all solutions in a flask and add the EDTA, solution has a purple color.
  • 2. Lead to boiling point, until the solution has a green color.
  • 3. Decrease a little the temperature (no less of 70 °C), set pH between 6.5-6.8 with KOH to 20%.
  • 4. Gauge to 1 liter.
  • 5. Leave the solution in a flask until the solution get a purple color.
  • 6. Remove the residue of reddish brown color by filtration.



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