How To Prepare MURASHIGE-SKOOG Medium

Murashige-Skoog (MS) medium is widely used for plant tissue culture because it has proven effective for growth promotion of both monocotyledons and dicotyledons.This medium is characterized by high  concentration of mineral salts, nitrate and ammonium which appears to be preffered by cells of some species.

Ingredients:

Macronutrients:

Ammonium nitrate (NH4NO3) 1,650 mg/l

Boric acid (H3BO3) 6.2 mg/l

Calcium chloride (CaCl2 · 2H2O) 440 mg/l

Cobalt chloride (CoCl2 · 6H2O) 0.025 mg/l

Magnesium sulfate (MgSO4 · 7H2O) 370 mg/l

Cupric sulfate (CuSO4 · 5H2O) 0.025 mg/l

Potassium phosphate (KH2PO4) 170 mg/l

Ferrous sulfate (FeSO4 · 7H2O) 27.8 mg/l

Potassium nitrate (KNO3) 1,900 mg/l

Manganese sulfate (MnSO4 · 4H2O) 22.3 mg/l

Potassium iodide (KI) 0.83 mg/l

Sodium molybdate (Na2MoO4 · 2H2O) 0.25 mg/l

Zinc sulfate (ZnSO4·7H2O) 8.6 mg/l

Na2EDTA · 2H2O 37.2 mg/l


Common organic additives:

i-Inositol 100 mg/l

Niacin 0.5 mg/l

Pyridoxine · HCl 0.5 mg/l

Thiamine · HCl 0.1 mg/l

IAA 1–30 mg/l

Kinetin 0.04–10 mg/l

Glycine (recrystallized) 2.0 g/l

Edamine S 1.0 g/l

Sucrose 20 g/l

Agar 10 g/l

REQUIREMNETS: Constituents of the MS medium, Erlenmeyer flasks (100, 250, 500 ml, 1 litre capacity), measuring cylinders (100, 1000 ml capacity), pipettes (1, 5, 10 ml), distilled or demineralized water, PH meter, 1.0 N NaOH, HCl or KOH and autoclave.


PROCEDURE:

1. Prepare macronutrients solution in 100 ml distilled water.

2. Prepare stock solution dilution chard.

3. Add macronutrients in 1-litre Erlenmeyer flask.

4. Add the other heat stable constituents (ex. sucrose, vitamins and harmones) and agar powder(if desired at a concentration of 0.8-1.0%).

5. Make the final volume of the medium by the addition of more distilled water.

6. Adjust pH of the medium to 5.7, using 0.1 N NaOH or 0.1 N HCl.

7. If solid medium is desired, agar is used.

8. Pour the medium into the desired culture vessels (15 ml in 125 * 150 mm culture tube and 50 ml in a 250 ml flask).

9. Plug the culture vessels with non-absorbent cotton wool wrapped in cheese-cloth or with any other suitable closure.

10.Transfer the culture vessels to appropriate baskets covered with aluminium foil to check wetting of plugs during autoclaving.

11.Transfer the baskets to autoclave.

12.Sterlize the medium by autoclaving at 121`c for the time period depending upon the volume of the medium in the vessel.

13.The medium is allowed to cool at room temperature and should be stored at 4 `c for future use.

Posted by:
Mitesh Jain.