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MEDIA PREPARATION FOR PLANT TISSUE CULTURE

Method · January 2017

DOI: 10.13140/RG.2.2.16632.44802

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Ali Talati
Shiraz University of Medical Sciences
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 MEDIA PREPARATION FOR PLANT TISSUE CULTURE

OBJECTIVE:

Media preparation is one of the primary and most essential steps in tissue culture. Media is
prepared based on the type of tissue being cultured. Most media differentiate from each other based
on the requirements of the growth of the specimen it supports.

 To understand the basic process of preparing media for plant tissue culture to promote.
 Growth and differentiation of tissues and acknowledge the basic requirements for the plant
Tissue culture.
 To prepare culture medium based on plant species requirements.
 To prepare stock solution consists of macronutrient, micronutrient and organic elements.
 To prepare culture medium by using aseptic technique.
MATERIALS:

 -Mutashige and Skoog (MS) medium (Powdered form)

 -800 ml of distilled water
 -30 g of sucrose
 -3.0 mg/ 1 BAP (As favourable hormones)
 -1.0 mg/ 1 BAP (As favourable hormones)
 -0.5 mg/1 NAA (As favourable hormones)
 -3.0 mg/1 Kinetin (As favourable hormones)
 _0.5 mg/1 IAA (As favourable hormones)
 -8 g agar

APPARATUS

 -Sodium hydroxide
 -Hydrochloride acid
 -Breaker and test tubes
 -Spatula
 -Volumetric flask
 -Autoclave
 -Sterile tube
 -Electronic balance
 -magnetic stirrer
 -PH meter
METHODS:

1. A packet of MS medium (in powdered form) is used for preparation of 1 litter medium.

2. 800 ml of distilled water is filled in a beaker.

3. MS powdered medium is slowly added into the beaker.

4. 30 g of sucrose is added.

5. PH is set at 5.8.

5. 8 g agar technical is added to the beaker.

6. Hormone is added.

7. The media is made up to 1 litre by using volumetric flask by adding 200 ml of distilled water.

9. The media is autoclaved.

10. The melting media is dispensed into sterile tubes. Each tube is labelled.

11. Each of the media listed below is prepared:

1. MS basal (without hormone)

2. 1.0 mg/1 BAP + 0.5 mg/1 NAA

3. 3.0 mg/1 BAP

4. ½MS + 3.0 mg/1 kinetin + 0.5mg/1 IAA

 A) Preparation of MS basal medium (without hormone)

1. 800 ml of distilled water is filled in a beaker.

2. 4.4 g of MS powdered medium (slowly added), 30 g of sucrose, and 8 g of agar technical are
added accordingly to the beaker.

3. The medium is made up to 1 litre using volumetric flask by adding 200 ml of distilled sterile
water.

4. The PH of medium is adjusted to 5 by using sodium hydroxide or hydrochloride acid.

5. The medium is then autoclaved.

6. The melting medium is dispended into sterile culture tubes for about one-third tubes’ height, in
laminar flow and each tube is labelled.

7. The condition of prepared medium is observed and used for culture a week later.

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