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Questions On Agar
    #590387 -

hello everyone, my friend is about to take his first steps into agar but first he asked me to help him clear up a few things........

1. could someone re-post the link that has a page of about 20 common agar recipes for cultivating higher fungi. i saw it while i was reading through every post that contained the phrase "agar" in the past year and now i cant find it for the life of me

2. did everyone order their petri's off the internet, i do not have a "science store" in my city and was wondering if there was anywhere else to get them (proably not, but its worth a try)

3. glass or plastic? i heard that glass is easy because you can PC them, but they do contam a little easier due to higher condensation....... whats your opinion?

4. will my friend be happy with a glovebox for a little while with his agar work? should he even attempt it without a flowhood? (he has little intrest in h202 culturing)

5. does PDYA = Potato Dextrose Yeast Agar???? i may be way off on this one, but i think i have the others pegged

6. where do you find dextrose and what is the household use for it? ED:: (just bodybuilding? would it be at a body supplement store? if its just corn sugar i will feel like a jack-ass) ::ED

7. after sealing your petri's with saran wrap (the non-name brand stuff) or tape or whatnot....do you put them in your incubator? or leave them in your glovebox?

well, that ended up being more than i thought, but they kept coming to me..... thanks alot


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"the weekend has landed all that exists now is clubs,drugs, pubs, and parties" - Human Traffic

Edited by ParticleMan (03/26/02 08:30 PM)

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Re: Questions On Agar [Re: ParticleMan]
    #590509 -

i'm in the same boat as you. i believe it's just plain white sugar, but could someone please clear this up? not 20 i don't think, but it's probably mostly the same.

Agar Formulas & Info
Amaranth Soy Agar 20 grams amaranth flour 20 grams soy flour 9.5 grams agar 500 mL distilled water Final pH: 6.0

Barley Malt Extract Agar 40 grams barley flour 2 grams malt extract 1 - 2 grams yeast extract (optional) 9.5 grams agar 500 mL distilled water Final pH: 6.0

EntheoGenesis No. 442 10 grams amaranth flour 10 grams brown rice flour 10 grams potato flour 10 grams soy flour 2 grams malted barley 9.5 grams agar 500 mL distilled water

Cornmeal Dextrose Agar 25 grams yellow cornmeal 2.5 grams dextrose 9.5 grams agar 500 mL distilled water Final pH: 6.0

Dr. Pollock's Modified Agar 10 grams dried dog food 10 grams amaranth flour 2 grams dextrose or malt extract 9.5 grams agar 500 mL distilled water Final pH is 6.2.

Malt Extract Agar 20 grams malt extract 100 mg potassium phosphate dibasic (K2HPO4) 100 mg calcium carbonate 9.5 grams agar 1 L distilled water
Oatmeal Neopeptone Agar 40 grams oatmeal or oat flour 2 grams neopeptone (optional) 9.5 grams agar 500 mL distilled water Final pH: 6.0

Modified Sabouraud's Medium 25 grams barley flour 5 grams dextrose 2 grams neopeptone (optional) 1 gram yeast extract 9.5 grams agar 500 mL distilled water Final pH: 5.8

Potato Dextrose Yeast Agar 250 grams of washed, unpeeled potatoes 10 grams dextrose 1.5 grams yeast 15 grams agar Slice potatoes 1/8 inch thick Rinse slices clean in cool tap water Final rinse with distilled water Cook in distilled water until tender Strain and set aside cooking water Rinse potatoes with distilled water Retain rinse water, discard potatoes Add distilled water to make 1 litre Bring liquid to slow boil Add dry ingredients
Cooking & Usage Instructions
?
The flask or bottle in which the medium is sterilized should never be more than 2/3 full in order to avoid boilover. ?

Plug the flask or bottle with non-absorbent cotton and cover with aluminum foil to help prevent entry of external contaminants. ?

Sterilize all formulas for 15 minutes at 15 psi. ?

Remove the medium as soon as the pressure reaches "0." Oversterilization or prolonged heating will change the composition of the medium. Sugars such as dextrose or malt can "caramelize," resulting in a medium that can inhibit mycelial growth and encourage sectors (mutations). Excessive heating of medium can also cause a drop in pH, resulting in a more acid medium. It is possible to destroy completely the gelling properties of agar by prolonged heating, and this destruction is hastened as the acidity increases. ?

For long-term health and maintenance of sacred strains, alternate any two of the above media. This will help prevent senescence, which can occur when cultures are grown on one medium only. Store strains at 35? - 40?F; transfer every six months. In this manner, sacred strains can be maintained for years. ? Never compromise the integrity of your strain library. Strains showing even a hint of senescence should be removed from the collection.





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Edited by Shaw (03/26/02 10:04 PM)

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Re: Questions On Agar [Re: ParticleMan]
    #590530 -

Good info above, table sugar is sucrose; a disaccharide in which one of the two saccharides is . . .glucose [AKA dextrose!]. The other, though, is fructose. Purified fructose. . .I don't know what effect it will have on fungi, but if you give someone a glucose IV during surgery you maintain life. . .if you give someone a fructose IV they die painfully.
It might work, I'd go with one of the other recipes [or eliminate the dextrose and go PYA] before adding table sugar to media.

I don't know, anyone ever substituted sucrose for dextrose?

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Re: Questions On Agar [Re: Suntzu]
    #590540 -

well i am about to give myself a fructose IV if i dont figure this out soon........no, not really......things are coming along ok, i am about to go mix up a batch using 15 ml corn syrup instead of 10 ml dextrose as a substitute. i would say i will let yall know how it goes.....but i can already tell you....i messed up, maybe not due to the sub. proably just because its my first try. BUT I'LL LET YOU KNOW :-)


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"the weekend has landed all that exists now is clubs,drugs, pubs, and parties" - Human Traffic

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Re: Questions On Agar [Re: ParticleMan]
    #590640 -

dang. Was hoping it was table sugar. Screwed now.
While were at it, whats potassium phosphate dibasic (K2HPO4). I have no idea whare it comes from or where to get it.
Gots the yeast, agar, potatoes,light malt extract, and brown rice flour. Now, to find a tek with those ingredients.


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Edited by Shaw (03/27/02 01:07 AM)

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Re: Questions On Agar [Re: Shaw]
    #590654 -

ok, apparently it requires a trip to the home brew shop. i should have figured that out.


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Re: Questions On Agar [Re: Shaw]
    #590721 -

well......i just found this in a document i looked at way in the beginning of my agar research and it said.........

PDA

10 ounces or 284 grams potatoes
3/4 ounce or 21.3 grams agar
1/4 ounce or 8 grams dextrose -- table sugar may be substituted <------ !!!!!!!

the page is.... http://www.mykoweb.com/articles/cultivation.html
it is listed at PDA method #2


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"the weekend has landed all that exists now is clubs,drugs, pubs, and parties" - Human Traffic

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Re: Questions On Agar [Re: ParticleMan]
    #590859 -

Good to know; Dextrose has probably been the 'standard' just because it's been the 'standard'.
It would be interesting to see comaprative growth on PFYA [pot. fructose yeast agar], PDYA, and PSYA [pot. sucrose yeast agar].
Hope it goes well--if your air quality is controlled pretty well and you sterilize/cool effectively, you should be good to go.

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Re: Questions On Agar [Re: ParticleMan]
    #590933 -

i wonder if honey is better than white sugar. I'll throw something together and see what happens. I also don't have any "non absorbent cotton" i'll just have to use the absorbent stuff and hope it works. i was really hoping that the agar would have alot less half assing involved, but oh well.


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Re: Questions On Agar [Re: Shaw]
    #591073 -

yeah, i hope we arent making something that has some difficulty to it impossible by half-assing it from the start.....i guess i just wanna try it out. if it doesnt work i will put in the extra effort to get everything exactly right.

i will be using.......

300 grams potatoes
20 grams agar
10 grams table sugar
2 grams yeast

now i read earlier that it does not matter if the yeast is "active" or not, when you PC its supposed to "deactivate" it. is this right????

Shaw,
I am cooking it in the quart jars that are sometimes used for grain, so i will just be using the poly-fil lids that people also use for grains........i dont think the cotton thing will matter.


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"the weekend has landed all that exists now is clubs,drugs, pubs, and parties" - Human Traffic

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Re: Questions On Agar [Re: ParticleMan]
    #591101 -

FOUND IT ! ! ! ! ! ! ! ! !

95. POTATO SUCROSE AGAR

Diced potatoes 300 g
Sucrose 30 g
Agar powder 15 g
Distilled water 1 L
Boil finely diced potatoes in 500 ml of water until thoroughly cooked; filter through cheesecloth and add water to filtrate to 1 L.
Add the agar and dissolve in the filtrate by heating. Sucrose is added prior to sterilization.

it doesnt mention yeast, but i am sure you could toss it in....... bonus

and just in case you need to see it for yourself.......
http://www.uv.es/cect/mlist.htm
its #95


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"the weekend has landed all that exists now is clubs,drugs, pubs, and parties" - Human Traffic

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Re: Questions On Agar [Re: ParticleMan]
    #591110 -

Dude, I'm impressed. Nice find.

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Re: Questions On Agar [Re: Suntzu]
    #591146 -

thanks, i hope it helps someone else, i decided to quit being a slacker and went and got some dextrose, malt extract, gypsum, and calcium carbonate at the store today.


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"the weekend has landed all that exists now is clubs,drugs, pubs, and parties" - Human Traffic

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Re: Questions On Agar [Re: ParticleMan]
    #591423 -

half assed it. This was my recipe.

2g nutritional yeast
5g brown rice flour
20g light malt extrace
15g agar

had several problems. the rice settled to the bottom. the polypropolene dishes didn't come sterilized as it was thought they would. the bag was cut in half for shipping, so had to resterilize the poured dishes. Didn't find it out till it was ready to pour.


also ran out of dishes. made half the recipie (1/2 leter) and filled all 20 plus 8 1/2 pint jars. could have done several more if the pourer was better at it. then, i threw em all back in the pc. the brf only got in the jars cause they were the last poured. Suran wrapped them, and there ready for innoculation.


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Edited by Shaw (03/27/02 10:32 PM)

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