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Registered: 09/19/01
Posts: 35
Loc: Troms?, Norway
PDA method.. Gonna give it a try!!!
    #505550 - 12/31/01 12:51 AM (21 years, 9 months ago)

So anyway, after two unsuccessful attempts with the BRF setup and shroomwizards casing, and 4 months, I'm pretty pissed and desperate to some have some fungi. Its got to work eventually, right.. so i hope.

so here's the PDA method I'm about to use... minus the yeast.

"Preparation Of Media"
PDA (Potato Dextrose Yeast Agar):
Wash 9 ounces of unpeeled potatoes and slice them 1/8" thick. Wash these several times in cool tap water until the water is clear. Drain these slices in a colander and rinse once in DISTILLED water. Cook the potato slices in 2 cups of DISTILLED water until tender. Strain the liquids through a piece of cloth in a strainer and collect the liquids. Rinse the potatoes 2 more times using 1 cup of DISTILLED water each time, add these rinse waters to the collected strained liquids and discard the potatoes.
Add enough DISTILLED water to the collected liquids to make 1 quart (4 cups). Bring the collected liquids to a boil in a pot and add 15 grams of agar, 15 grams of dextrose and 1.5 grams of nutritional yeast (you can purchase all three of these items in about any Health Food store). The agar should be added slowly to avoid boiling over. While the liquid is still hot, pour it into 1/2 pint canning jars until about 1/4 full. Sterilize these jars of agar medium in boiling water as described earlier under "Sterilization".
PDY (Potato Dextrose Yeast Broth): PDY Broth is made in exactly the same manner as PDA except the agar is omitted. One quart canning jars are filled 1/2 full of this broth and sterilized as described earlier.
"Starting The Culture"
A single suspension syringe contains millions of spores and any one of these will germinate in an agar medium to form a mycelium (the root system of the mushroom plant).
Using the Pure Culture Technique, remove the lid carefully from one of the 1/2 pint jars of agar you prepared (and watched for 3 days). Hold the suspension syringe at a sharp angle over the open jar and inject 2-3cc's of Suspension into the agar jar and replace the lid. As a rule of thumb, if you can see any of the Suspension on the agar, that's enough. There is no reason to waste a whole syringe on just one agar jar.
Within 3 to 5 days you should see what resembles mold growing across the surface of the agar. What you are looking for is a snow white growth, this is the mycelium of the Psilocybe Cubensis mushroom plant. If any other color appears it is a foreign fungus and isn't wanted. If the only color that appears is snow white, then consider yourself lucky, but if different colors appear you have to use the "Pure Culture Technique" to snag a small piece of the pure white mycelium from the contaminated agar jar (I use flame sterilized tweezers points) and transfer it to a new agar jar where it can grow uncontaminated.
When the mycelium has almost covered the surface of the agar in the jar (3 to 5 days) it is time for inoculation into the 1/2 filled broth jars you have previously prepared and watched for 3 days.
"Inoculation Into Broth"

Now is the time to transfer the mycelium growth from the agar jar into the 1/2 filled one quart jars of PDA broth. Using the "Pure Culture Technique", transfer a small amount of the pure white mycelium (about the size of a match head) from the agar medium into one of the jars of prepared broth that you have prepared and observed for 3 days for foreign growth. Tighten the lid and shake the jar, this assures that the liquid broth has plenty of oxygen distributed throughout and also helps break up the mycelium for faster growth. Loosen the lid on the jar slightly and set on a shelf at room temperature. Continue with each jar of broth until all prepared jars have been inoculated. Every 2 days tighten the lid on the jar(s), shake to distribute oxygen and break up the growing mycelium, reloosen lid(s) and put back on shelf. In the beginning (the first few days) you will see what looks like white clouds in the liquid broth, this is the young mycelium plant. Within 10 to 12 days this will turn into a thriving mass of mycelium root system that is ready for harvest. All you do at this time is open the jar(s) and pour into a colander to drain. What you should have is 2 to 4 ounces of white mycelium per jar (wet weight). Since this mycelium is the same chemical composition as the mushroom fruit itself, you can prepare it in the same way you would prepare the mushroom.

So anyway, I've had two unsuccessful attempts at using the BRF mmtek method.. basically slow growth and green mold seem to work better than growing mycellium. if i wanted to grow mold i'd just leave a loaf of bread in a dark warm place. but thats not the point.. this time it should work.. I've found a tek to make a at home/poor dirty bastard glove box that should work pretty good to keep the contams away while i innoculate with a spore print.. I plan on doing part of my setup like the tek says, putting some in the few petri dishes i have, and putting the rest in some pint caning jars I still have laying around using them as petri dishes basically... if all goes well hopefully I'll have some serious mycellium growing without contams.. lets hope, right? after the mycellium grows (cross my fingers), I plan on doing some like the tek says (the innoculation broth), and transferring some into a BRF setup, and possibily if i can get a good cheap pressure cooker using rye berries as a substrate for my spawn... if anyone has any feedback, input , or questions let me know..... I really would love some feedback!!!!!

peace out.....

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Registered: 07/26/01
Posts: 556
Re: PDA method.. Gonna give it a try!!! [Re: djveiox]
    #505708 - 12/31/01 07:51 AM (21 years, 9 months ago)

i never read all that but in the last paragraph this is what i got ... you failed at the brf/verm jars because of green mold and you think that you're not going to get get mold on pda? hahahah...

-SumGuy :cool:

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