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1. Preparing Potato Extract
Place potatoes in a saucepan with approximately 1.2 liters of tap water and boil on a stove for 1 hour.
Remove saucepan from heat and let liquid cool until it can be comfortably handled (about 10 minutes).
Pour liquid potato extract through a strainer or cheesecloth and collect it in another container.
2. Preparing the Medium
Add dextrose and agar to 1 liter of the potato extract water.
Test the pH of the solution with pH test paper or a pH meter.
If the pH is above 6.5, adjust it down by adding one drop of the hydrochloric acid solution at a time and retesting.
If the pH is below 6, adjust it up by adding one drop of the sodium hydroxide solution at a time and retesting.
3. Sterilization
Thoroughly mix the medium and divide it into smaller batches of 250 mL poured into the four 500-mL flasks (Note: each flask must be no more than half full to avoid boil over).
Cover each flask with a piece of loosely crimped aluminum foil, so the foil will not fall off, but does not make a tight seal.
Place the flasks in a pressure cooker.
Operate the pressure cooker in accordance with the manufacturer's instructions to sterilize the PDA at 121 degrees C (250 degrees F) at a pressure of 103,421 Pascals(15 PSI) for 15 minutes.
Shut off the pressure cooker and let it return to atmospheric pressure, then remove the flasks and let them cool until they can be handled.
4. Pouring the Medium
Line up the sterile petri dishes on a flat surface.
Once the flasks have cooled to handling temperature, but before the medium begins to solidify, grasp a flask with one hand and remove and set aside its foil lid with the other hand.
For each petri dish, lift the lid with one hand, quickly fill halfway with liquid PDA, and replace the lid.
Repeat Step 3 above until all petri dished are filled.
If there is any PDA left over, loosely recover the flask with its original foil lid, let it cool and solidify, tighten the foil lid and store in a refrigerator.
References: http://www.ehow.com/how_5872701_make-potato-dextrose-agar.html |
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