DATE:

   NAME:

   The theme: Preparation of Tissue Culture medium M-S

                     Steps in the preparation of 1 L of medium

   

   1.    Weigh 6,5 g of agar and put it into 300 ml deionised, distilled water in SIMAX bottle,
   stir and melt it in autoclave.

   2.    Put 500 ml of deionised, distilled water into an Erlenmeyer bottle.

   3.    Add the appropriate amount of the macro elements stocks (100 ml), microelements stocks
   (10 ml) and chelatonized iron stock (5 ml).

   4.    Add the vitamins  (1 ml freeze concentrate).

   5.    Weigh 100 mg of inositol.

   6.    Weigh 20 g of sucrose.

   7.    Add plant growth regulators and other components, if necessary.

   8.    Adjust the pH using 0,1 M KOH or 0,1 M HCl. The pH should be 5,7.

   9.    Add the solution into SIMAX bottle containing melted agar; adjust the final volume  (1L)
   with deionised, distilled water.

   10. Thoroughly mix and distribute the medium into the tissue culture vessels (40 ml into each
   vessel).

   11.Cap the vessels.

   12. Autoclave for 20 min at 121°C. If a large quantity is being autoclaved, longer times may
   be necessary.

   13. For short-term storage keep media at room temperature, for prolonged storage keep media in
   the refrigerator.