CM-0015
Waris-H + 3V

55,00 €

Información del Medio de Cultivo

Waris-H + additional vitamins

(McFadden G.I & Melkonian M., 1986)

#	Component	Stock solution Addition per 1 Litre stock solution	Addition per 1 Litre culture medium
1	KNO₃	100.00 g	1 ml
2	MgSO₄ ^. 7 H₂O	20.00 g	1 ml
3	(NH₄)₂HPO₄	20.00 g	1 ml
4	Ca(NO₃)₂ ^. 4 H₂O	100.00 g	1 ml
5	HEPES	238.31 g	1 ml
6	P-II Metals		1 ml
7	Fe-EDTA		1ml
8	Vitamins		3 ml
9	Soil extract		10 ml

Preparation of PII Metals Solution

#	Component	Stock solution Addition per 1 Litre stock solution
1	EDTA (Titriplex III)*	3.00 g
2	H₃BO₃	1.14 g
3	MnCl₂ ^. 4 H₂O	0.144 g
4	ZnSO₄ ^. 7 H₂O	0.021 g
5	CoCl₂ ^. 6 H₂O	0.004 g

Preparation of Fe-EDTA Solution

#	Component	Stock solution concentration Addition per 1 Litre stock solution
1	EDTA (Titriplex II)*	5.22 g
2	FeSO₄ ^. 7 H₂O	4.98 g
3	1 N KOH	54.00 ml

EDTA (Titriplex II) and FeSO₄ x 7 H₂O is heated for 30 min (100^oC); KOH is added to the cooled mixture

Preparation of Vitamins Solution

#	Vitamin	Stock solution concentration Addition per 1 Litre stock solution
1	Vitamin B₁₂	0.20 mg
2	Biotin	1.00 mg
3	Thiamine-HCl	100.00 mg
4	Niacinamide	0.10 mg

Preparation of Soil Extract

10g of garden-soil is mixed with 120 ml bidistilled water and boiled for 10 minutes. Afterwards it is centrifuged for 10 minutes (low speed), and the supernatant is filtered through a series of membrane filters from 1.2 µm – 0.1 µm pore size. The remaining filtrate is adjusted to 100 ml with bidistilled water. Aliquots of 10 ml are stored frozen. The soil should not be recently fertilized and should not contain too much humus.

Preparation of Culture Solution

Add 1 ml of stock solutions 1-8 to 1000 ml of bidistilled water. Add 10 ml of thawed soil extract (= stock solution 9), adjust the pH to 7.0 and autoclave.

* Titriplex II - Ethylenediaminetetraacetic acid - C₁₀H₁₆N₂O₈

* Titriplex III - Ethylenediaminetetraacetic acid disodium salt dihydrate - C₁₀H₁₄N₂Na₂O₈x 2 H₂O