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Homemade PDA+ Agar Plates Illustrated


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#1 Hippie3

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Posted 29 January 2008 - 08:28 PM


Homemade PDA+ Agar Plates Illustrated

Materials Needed

300-350 grams potato (1 large potato)
14 grams agar (from biological supply, health food store or Asian food market)
10 grams Dextrose or other sugar
2 grams nutritional yeast (optional)*
2 grams amaranth flour (optional)*
2 one quart jars, bottles or flasks
50 sterile plates (15x100)
*note: for long term storage of cultures I highly recommend adding at least one of these

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Scrub the potato but do not peel. Slice into thin slices. Boil potato in 1.2 liters of filtered or distilled water for 45 min to 1 hour. Drain the potatoes, reserving the hot liquid. Bring to 1 liter with additional boiling water as needed.

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Place the potato broth (1 Liter), the agar, sugar and optionally the yeast and flour in a bowl and mix thoroughly (a wire whisk works well, but stir, do not beat). You need to do this while the broth is hot so that the agar will melt.

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Pour half of the mixture into each container. Fill to only 1/2 of the vessel's capacity. Plug the mouths of the bottles or flasks with cotton or polyfill and cover with aluminum foil. Bring the water in your pressure canner (PC) to a boil. Put jars into PC. Cover and allow steam to vent for 5 min. Close petcock and "cook" at 15 psi (250?F) for 15 minutes. Try to not let the temp rise above 250?F (15 psi) as you can caramelize the sugar at higher temps.

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After the 15 minutes, turn off the heat and allow the pressure to return to normal gradually or with gentle venting (about 30 minutes). Having the containers half-full will prevent boil-over if you are careful about venting.

Get your plates, some gloves, and a HEPA mask. Put on the mask and gloves and sanitize your hands and arms with whatever you want. I use cheap vodka.

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Put your plates out on a clean surface.

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Remove your agar as soon as the pressure reaches zero but it is still hot. Using a hot mitt, remove the jar from your PC. Swirl the agar to make sure it is thoroughly mixed.

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Open your jar of agar. Lift the lid on each plate and pour in agar just 'til it covers the bottom. Replace lid on plate. This is a two handed operation and the lid should just be tilted up during the pour and then quickly replaced. Continue to swirl the agar as you are pouring the plates.

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(I had to go back later and take this pic, but you get the idea.)

500 mL of agar will do 25 or so plates. If you only pour 25 plates, the other jar of agar can be saved in the fridge. When you are ready to use it, look for absence of growth. If its clean, just pop it in your PC and cook it as above.

After you pour, condensation will form on the lids, but don't worry.

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Allow the plates to cool for about an hour. Label plate lids with contents (PDA+) and date but leave room for more info. Turn them over, stack and resleeve.

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Plates can be stored like this for months in a refrigerator. If stored, be sure to check for mold or bacterial contamination before use.

Plates can be used for live tissue culture, mycelium culture, germinating spores, etc.

  • souravhajra5 likes this

#2 hobolicious

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Posted 29 January 2008 - 09:12 PM

Sweet guide. Can you PC the plastic plates after they're filled? FOAF was hoping to seal the edges and PC them before using them in a glovebox.

#3 Hippie3

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Posted 29 January 2008 - 09:13 PM

nope, they'd melt/boil over.
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#4 Scroto Baggins

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Posted 29 January 2008 - 09:21 PM

:bow:Right on Hip! This is exactly the kind material that is invaluable to a budding Mycophile like myself. I am definitely going to use this recipe when I do my first agar attempt to try and clean up a villarrealii print.

One question though. Can this mixture be sterilized through just steaming or perhaps using Fractional Sterilization?

#5 Hippie3

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Posted 29 January 2008 - 09:23 PM

i would think so, but never personally tried.

#6 hobolicious

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Posted 29 January 2008 - 10:01 PM

Wait, the agar would melt, or the plastic? So your best bet is to pour in a glovebox or in front of a hood? Also, what about guar/xanthan? Could that be PC'd, or at least steamed at < 150 degrees? I'm new to agar and I'm looking to minimize contams. You know the old saying:

"A smart man learns from his mistakes. A wise man learns from the mistakes of others."

#7 Hippie3

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Posted 29 January 2008 - 10:03 PM

So your best bet is to pour in a glovebox or in front of a hood?


correct.


guar = trash

I'm new to agar and I'm looking to minimize contams.

just follow the directions.

Edited by Sidestreet, 14 August 2015 - 10:44 AM.
broken link

  • usagolden likes this

#8 gsmith1981

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Posted 29 January 2008 - 10:21 PM

:cool:

#9 OG Royal Grower

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Posted 30 January 2008 - 04:52 AM

way to save the day hip:eusa_clap




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