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OfflineSick_Boy
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1st time Agar + Pitri Dish - Help
    #14327115 - 04/21/11 10:31 AM (12 years, 10 months ago)

So today I picked up some pitri dishes and all the ingrediants I need to make PDA

the recipe i got from Fastfreds media cookbook
Potato Dextrose Yeast (PDY)

200 g Potato, infusion from [dilute to 1L total] (im using potatoe flake)

20 g Dextrose
  2 g Yeast extract
20 g Agar 
  1 L Distilled water (dH2O



My question, After I mix this all up. Do I heat up the liquid and pour it into the pitri dishes and then PC the pitri dishes?  My pitri dishes are plastic not pyrex?  Please let me know as im eager to try this out.

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InvisibleEvilMushroom666
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Re: 1st time Agar + Pitri Dish - Help [Re: Sick_Boy]
    #14327156 - 04/21/11 10:41 AM (12 years, 10 months ago)

You are going to want to mix the ingredients in a container that you
will PC and then use to pour into your petri dish's under sterile
conditions.

Here are some write ups I did when I was having my first crack at
agar that might help you out:

Agar Setup(Costs and Materials)
https://www.shroomery.org/forums/showflat.php/Number/12675513

Pouring Malt Extract Agar
https://www.shroomery.org/forums/showflat.php/Number/12748848

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OfflineSick_Boy
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Re: 1st time Agar + Pitri Dish - Help [Re: EvilMushroom666]
    #14327190 - 04/21/11 10:52 AM (12 years, 10 months ago)

Thanks

When using a spore print can i scrape a sample directly onto the agar or do i need to rehydrate it first?

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InvisibleEvilMushroom666
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Re: 1st time Agar + Pitri Dish - Help [Re: Sick_Boy]
    #14327211 - 04/21/11 10:57 AM (12 years, 10 months ago)

Use a sterile inoculation loop to swipe your spores onto the agar,
of course under sterile conditions.

If the spores do not readily germinate you can try making your agar
on the wet side(cut back on the amount of agar in the recipe).

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OfflineSick_Boy
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Re: 1st time Agar + Pitri Dish - Help [Re: EvilMushroom666]
    #14327327 - 04/21/11 11:36 AM (12 years, 10 months ago)

thanks for your help evil.

So i modified the recipe, let me know if this makes sense.  This recipe is for 1L.  Im using a 500ml jar as my bottle. so I used half a cup of water. and rounded it up for 250ml

5g Dextrose
5g agar
.5 yeast
7.5g potatoe flake

right now its PC'ing, 90 min or 60?

thx

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InvisibleEvilMushroom666
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Re: 1st time Agar + Pitri Dish - Help [Re: Sick_Boy]
    #14327343 - 04/21/11 11:40 AM (12 years, 10 months ago)

Iv never used the recipe so I am not sure, I mostly use MEA,

Per 500ML water:

10G agar
10G Light Malt Extract


I usually PC my agar for 45 minutes at 15PSI.

The bottle you are using has a filter? Tyvek, polyfil or synthetic
filter disc?

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OfflineSick_Boy
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Re: 1st time Agar + Pitri Dish - Help [Re: EvilMushroom666]
    #14327467 - 04/21/11 12:13 PM (12 years, 10 months ago)

The bottle im using is a jar, which does have a hole in the jar.  I have the bottle wrapped in tinfoil right now but there is nothing covering the hole.

My plan is to leave it in the PC to cool down. then open the PC and place the jar with foil still on it into my still air box and work from there.

is this ok?

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InvisibleEvilMushroom666
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Re: 1st time Agar + Pitri Dish - Help [Re: Sick_Boy]
    #14327508 - 04/21/11 12:23 PM (12 years, 10 months ago)

You need to provide a filtered port in your container so that as your
PC cools and draws in un clean air it does not contaminate the agar.

I use polyfill pulled through a hole in the lid of a whiskey bottle
for my filter. Some use tyvek circle or synthetic filter discs.

Also you want to pour agar while it is still warm. What I do is wait
for my PC to cool down so I can touch the outside then remove the
bottle and place in front of my flow hood to cool some more. When I
can touch the glass bottle without it burning my hands I know it is
ready to pour.

If you let the agar cool completely it will harden and need to be heated
back up to become liquid again.

P.S Whiskey and liquor bottles will be easier to pour from then jars.

Edited by EvilMushroom666 (04/21/11 12:29 PM)

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OfflineSick_Boy
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Re: 1st time Agar + Pitri Dish - Help [Re: EvilMushroom666]
    #14327574 - 04/21/11 12:42 PM (12 years, 10 months ago)

Thanks for the advice, that is a shame because I have polyfil I could have used and just didnt realize I was risking contamination.

I guess Ill have to just try my luck, and hope it works out.

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InvisibleEvilMushroom666
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Re: 1st time Agar + Pitri Dish - Help [Re: Sick_Boy]
    #14327591 - 04/21/11 12:45 PM (12 years, 10 months ago)

No problem and good luck!

If the petri dish's you are using are expensive($10 a sleeve) you could
always make up another batch using a proper bottle.

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Re: 1st time Agar + Pitri Dish - Help [Re: EvilMushroom666]
    #14327628 - 04/21/11 12:56 PM (12 years, 10 months ago)

I got a good deal so im going to give it a shot. I do have the tin foil wrapped very tightly around the lid with an elastic band double wrapped holding the tin foil down tightly.  The jar is sitting in my glove box right now. after its been well cleaned.

Will see. and thanks for all the help!

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OfflineSick_Boy
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Re: 1st time Agar + Pitri Dish - Help [Re: Sick_Boy]
    #14328006 - 04/21/11 02:36 PM (12 years, 10 months ago)

Question guys,

Should I re-hydrate my spores before dropping them on the Agar pitri dishes? I know the agar wont be very wet once its done so?

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Offlinesteelmonkey
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Re: 1st time Agar + Pitri Dish - Help [Re: Sick_Boy]
    #14328515 - 04/21/11 04:11 PM (12 years, 10 months ago)

Just make your agar a little on the wet side as Evil stated and they will hydrate themselves good luck on your first agar work i'm glad to see your trying it


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OfflineiSHACKA
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Re: 1st time Agar + Pitri Dish - Help [Re: steelmonkey]
    #14328636 - 04/21/11 04:34 PM (12 years, 10 months ago)

DELETE


--------------------
Pain is just weakness leaving the body

Edited by iSHACKA (04/22/11 12:03 PM)

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Offlinesteelmonkey
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Re: 1st time Agar + Pitri Dish - Help [Re: iSHACKA]
    #14328817 - 04/21/11 05:08 PM (12 years, 10 months ago)

Are you waiting until they are completely cooled before putting the lids on? I like to leave the lids open a crack in the glove box till they've cooled also how are you storing them? where?


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OfflineiSHACKA
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Re: 1st time Agar + Pitri Dish - Help [Re: steelmonkey]
    #14329066 - 04/21/11 05:59 PM (12 years, 10 months ago)

OOPPS


--------------------
Pain is just weakness leaving the body

Edited by iSHACKA (04/22/11 12:03 PM)

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Offlinesteelmonkey
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Re: 1st time Agar + Pitri Dish - Help [Re: iSHACKA]
    #14329115 - 04/21/11 06:08 PM (12 years, 10 months ago)

That all sounds great and its correct you should have the lid on asp try leaving the lid on but cracked just a touch in the glove box maybe put a peice of alcohol sprayed paper towel over them


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