Medios de cultivo Medios de cultivo

CM-0014
Waris-H

55,00 €

    Waris-H Medium

    (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

     

    1 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 (100oC); 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  - C10H14N2Na2O8 x 2 H2O