Preparation of Media For Culture-PDA, PDB & NA Recipes

PREPARATION OF MEDIA FOR CULTURE:
Media play an important role not only for the nutrition and growth of the pathogen but also in identification through growth pattern on media. Fungi and Bacteria need synthetic media enriched with essential element for the growth. In general different types of media are prepared to culture fungi and bacteria but Potato dextrose agar (PDA) for fungi and Nutrient agar for bacteria are being used commonly. 
RECIPES OF DIFFERENT MEDIA:
POTATO DEXTROSE AGAR:

Potato Starch 250 mL/15g

Dextrose-15g
Agar -Agar 15g

Distilled water 750 mL/1000mL

PROCEDURE TO PREPARE POTATO DEXTROSE AGAR MATERIAL:
Petri dish, Flask, test tubes, Agar-agar, Dextrose, Potato starch, Water
PROCEDURE:
First of all take potatoes to extract potato starch. Peel the potatoes and cut into small pieces. Preferably 200 g of potato pieces are weighed, washed quickly in running water, placed in 1l liter of water and boiled for nearly 1 hour till a mash is formed. The mash is then squeezed through double layer of muslin cloth to obtain as much of the pulp as possible. Discard the potato pieces. Alternatively potato starch commercially available in the markets can also be used. Take the potato starch 250 mL, add dextrose (15g) into it and slightly shake it so that dextrose completely mixes into starch. Now slowly add agar-agar (15g) into this and continue shaking until the whole contents get mixed thoroughly. Now add the distilled water (750 mL) to make up the volume as per requirement. Plug in the flask and autoclave. After autoclaving, medium is ready to pour into Petri dishes.

POTATO DEXTROSE BROTH:
Potato starch 250ml/15g
Dextrose  15g
Distilled water 750ml/1000ml

PROCEDURE TO PREPARE POTATO DEXTROSE BROTH MATERIAL:
Petri dish, Flask, test tubes, Dextrose, Potato starch, Water
PROCEDURE:

First of all take potatoes to extract potato starch. Peel the potatoes and cut into small pieces. Preferably 200 g of potato pieces are weighed, washed quickly in running water, placed in 11 liter of water and boiled for nearly 1 hour till a mash is formed. The mash is then squeezed through double layer of muslin cloth to obtain as much of the pulp as possible. Discard the potato pieces. Alternatively potato starch commercially available in the markets can also be used. Take the potato starch 250 mL, add dextrose (15g) into it and slightly shake it so that dextrose completely mixes into starch. Now add the distilled water (750 mL) to make up the volume as per requirement. Plug in the flask and autoclave. After autoclaving, medium is ready to pour into Petri dishes.

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Written By:

Dr.Qaiser Shakeel

Tahir Mahmood

Department of Plant Pathology-Agriculture

The Islamia University of Bahwalpur-Pakistan.