37 replies to this topic

[Hippie3]

you're on the right track,
it will be contaminated at first
so you'll need to cut out good sections for transfer,
once you get growth.
hope you post pix of the process
as we need more examples to share

[MurCurY]

i soaked my specimen in sterile "pressurecooked" water overnight. It actually looked like a mushroom cap again. I broke the cap into a few smaller pieces and plated them 2 days ago. So far...one plate has a white spot growing on the piece of cap...i'm hoping it's mycellium and not something funky. The other 2 plates...nothing so far.

[MurCurY]

update

The one piece that had a fuzzy spot on it....yesterday i looked and there were 3 fuzzy spots!!

i took jars i made up for another project and transferred a piece of mycellium from each of the 3 spots to new jars. The fuzzy spots appeared around the outer edge of the cap...so i'm assuming i may have gotten lucky and got a few spores to germinate.

Since I failed to mention this.....the cap i'm working with is from college....from 1999!! And it has never been in a refrigerator. Just bagged within a bag. I'll take pics later when i charge my batteries.

[shobimono]

Here's a couple recipes I had saved from somewhere.

These recipes are designed for agar maintenance of Psilocybe species. All media have a near-neutral pH and should be sterilized for twenty to thirty minutes at 15 psi pressure. 500 ml of medium will pour around 20 petri dishes. Using a different medium from time to time will prevent the culture getting used to one medium.

Malt Extract Agar (MEA)
10 grams light malt extract
9 grams agar agar
500 ml potable or distilled water

Potato Dextrose (Yeast-extract) Agar (PD(Y)A)
broth from boiling 150 grams sliced potatoes
in 500ml water for 30 minutes(add water to 500ml)
(You can use 5 g of instant potato flakes instead)
9 g agar agar
7 grams dextrose( = glucose, or 10 ml honey or corn syrup )
1 gram brewers yeast or yeast-extract (optional)

Amaranth Soy Agar
20 grams amaranth flour
20 grams soy flour
9 grams agar agar
500 ml potable or distilled water

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 grams agar agar
500 ml potable or distilled water

Oatmeal Neopeptone Agar
40 grams oatmeal or oat flour
2 grams neopeptone (optional)
9 grams agar agar
500 ml potable or distilled water

Modified Sabouraud's Medium
25 grams barley flour
5 grams dextrose
2 grams neopeptone (optional)
1 gram yeast extract
9 grams agar agar
500 ml potable or distilled water

Cornmeal Dextrose Agar
25 grams yellow cornmeal
3 grams dextrose
9 grams agar agar
500 ml potable or distilled water

Barley Malt Extract Agar
40 grams barley flour
2 grams malt extract
1 gram yeast extract (optional)
9 grams agar agar
500 ml potable or distilled water

Dr. Pollock's Modified Agar
10 grams dried dog food (ground to flour)
10 grams amaranth flour
2 grams dextrose or malt extract
9 grams agar agar
500 ml potable or distilled water

[MurCurY]

i've actually decided to use MEA since alot of people here use that. Besides the carmelization of some of the malt extract the spores seem to like it better then the home made PDA i made.....also though....the bacteria and other spores also like it.....

[golly]

Interesting project,,,did u treat the cap in any way to lower the risk of compeditors growing,,,also do u think that it's spores germinating or the tissue regenerating,,,? Hope it works out for ya...

[MurCurY]

from a single cap. I soaked overnight in pc'ed water...with VERY little water in the jar...just enough to hydrate the cap. Then i put it on agar...no other prepping. I figure if anything grew from it it's not a complete waste. I was telling the professor at school...and he doubted it would work at all. and i quote..."if anything you might get a couple of spores to germinate"
I'm not really sure if it's spores or if it's the tissue regenerating. Only one of the three jars have any kind of activity. Also...the cap is about 6 years old...and it wasn't stored in the most favorable conditions. I'll keep this thread up to date. Like i said...as soon as the batteries are charged i'll be taking pics.

[Hippie3]

murcury-
how did this finally turn out, btw ?

[Hippie3]

[editorial note: i merged an older but relevant thread by sandman with
murcury's thread to get the info in one place.
archive material ]

[FLcubiefan]

A foaf did that karo blender trick and it worked. His jar didn't get a thick mat of mycelium but when he shook it up and then used it to innoc it grew well. He didn't sterilize the needle though so the jars all got contams.

Hmmmm.. The magic bullet....

[Lazlo]

By the way, if you want to make your dry tissue cloning life more simple; hydrate the tissue and apply enough to a dish to surfice. After you've applied the hydrated tissue to agar, cover the tissue with another layer of hot agar immediately. It eventually grows right through to the top layer cleanly and can be transfered to a new dish of agar for further use....

Hot agar technique provided from Stephen L. Peele of FMRC...

[Hippie3]

i still wanna see someone try that
with beeswax instead of agar...

[MurCurY]

murcury-
how did this finally turn out, btw ?

grew some good mold...that's about it. i'll try again...still have some of the piece of mush to play with.

[Hippie3]

bummer.
try, try again.

[Lazlo]

Have you tried the hot agar pour as mentioned above? It takes time for the tissue to revive and to colonize through to the top layer of newly poured agar. But, it's very simple.

[MurCurY]

not yet...will give it a go when i get some time to play with this.

[jestah]

Hi

can you use any dryed mushroom? Im wanting to try my hand at an edible could i use a dryed shitiki from a pack?

thanks
jestah

[Lazlo]

Sure. It's tricky, but i've been using the hot agar pour and it seems very good. I've got some tissue from a dried out piece of cardboard that was used for a clone going now. It's been a couple of weeks and it appears that it's starting to revive. I'll post some pictures of the process if it works like my previous attempts.:eusa_pray