Potato Dextrose Agar (PDA)!

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hamloaf
Pork Prophet

Registered: 12/23/09
Posts: 29,889
Loc: Greasy forked truth typer

Potato Dextrose Agar (PDA)!

2
#12936745 - 07/22/10 07:40 PM (16 years, 3 months ago)

Make enough Potato Dextrose Agar to fill 25 3 sectioned petri dishes from Cynmar.
http://www.cynmar.com/item_detail.aspx?ItemCode=15023320

Supplies Needed:

Dried, instant potato flakes.
Powdered food grade agar agar (Mycologist grade agar agar, best).
Dextrose.
A liquor/whiskey bottle (the longer the neck of the bottle, the better).
Painter's cover alls tyvek, cut into 2 3by3 inch squares.
A funnel.
Distilled water.
A screw gun or drill with a quarter inch drill bit.
Digital scales.
1 teaspoon or tablespoon.
1 4by4 inch square sheet of foil.
1 pair of scissors.
1 measuring cup that weighs in milliliters.

Preparation of Your Liquor/Whiskey Bottle to an Agar Container.

First, prepare your liquor/whiskey bottle by consuming or giving the liquor to your friends.
Next, wash the bottle with dish soap, removing the sticker from the bottle.

Take your drill out.
Pop a quarter inch hole through the center of the bottle's cap.

Screw the cap onto the bottle to drill the hole.
With the cap screwed to the bottle, hold the bottle with one hand.
Drill the hole with the other hand.

Scrape away the shavings, smoothing out your freshly drilled hole.
Remove the shavings from inside your bottle and....Done!
You are ready to mix your agar ingredients into the liquor/whiskey bottle.

Portioning and Mixing Your Agar:

7.5 grams of dried potato flakes.
13.5 grams of powdered agar agar.
10.5 grams of dextrose.
750ml of distilled water.

Place funnel through the top of the bottle.

Weigh each of the ingredients out.
Add your ingredients into your bottle through the funnel.

7.5 grams of dehydrated potato flakes.

13.5 grams of powdered agar agar.

10.5 grams of dextrose.

Once you have placed all of your ingredients into the bottle measure out 750ml of distilled water in your measuring cup.

750ml of distilled water.

Pour half of your water into the bottle through your funnel.
Give your water and ingredients a good stir to mix the ingredients through out the water.
You are generally going to want to give you agar a stir between each step.
This will keep all the particles in suspension evenly through out your agar mix.

Add the rest of your distilled water and stir again.

Take your 2 3by3 inch squares of tyvek and place them on top of one another.
Place the doubled up tyvek squares over the bottle's top between the threads of the bottle and the cap.

Give your the contents of your bottle another good stir and.....Done!
You are now ready to PC your agar bottle and it's contents.

Preparing Your Agar Bottle and Contents for Sterilization in Your PC.

After you have prepared your bottle and mixed your agar your bottle and your bottle's contents will need to be sterilized.

Take your 4by4 inch square of aluminum foil wrap it firmly over the top of your bottle's lid and tyvek.
Be sure all of your tyvek is covered by the foil.

This will protect your tyvek filter from becoming wet caused by the dripping condensation from the lid during the PC cycle.

Take your bottle and contents and place it into your PC.
I fill the PC with water until the water level matches the level of the contents in the bottle.
This way the bottle, contents, PC, and water will all heat up at the same rate.

Turn your stove on high.
Bringing the water to a boil.
Once the water is at a full boil remove the pressure regulator(jiggler) and secure the lid to your PC.

Wait until steam is shooting out the vent pipe for 5 full minutes then place the pressure regulator(jiggler) on top of the vent pipe to build pressure.

Once your steam gauge reads 15 PSI adjust your heat on the stove to hold the temp of your PC, at 15 PSI for 90 minutes, autoclave at 15 PSI for 35-45 minutes.

Let PC cool for 2 hours before pouring your agar.
Pour your agar in front of a flowhood or in a glove box.

Once your agar is poured into the petri dishes, place them back into the bag they came in.
Let agar cool to room temps in front of a flowhood or glove-box.

Place in the fridge for storage (if needed).
HL

--------------------

How I Get Stuff done. - My Reference Guide. - My Grows.

Edited by LiquidMyce (07/23/10 12:39 AM)

Extras:

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