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Mushrooms, Mycology and Psychedelics > Mushroom Cultivation
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First AGAR atempt log...
    #5702019 -

Hello again everyone...Well, I figured it was high time i learn how to isolate strains on agar. So after doing much reading i have decided to go full blown.. With the help of The Mushroom Cultivator by Stamets. and a variety of other sources (including this forum!!!) I have begun my journey.

I am going to start with two strains. HAWAIIAN and PF. (chosen randomly)
As I am writing I am listening to the soothing hiss of my pressure cooker at it sterilizes my agar mix in a flask and 4 test tubes ( to make slants for later)

I have used a mixture of:
10g agar powder
10g malt extract
2.5g peptone
500 mL of distilled water.

Mixed well in flask then put a little in each test tube and covered all with foil and here we are.. now i am waiting for the 30 min timer to go off and i will stop the cooker and wait for it to cool off a little so that i can handle the flask and go pour my dishes in my glove box... (wish me luck folks) hope everything goes well...

MJ


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All posts from this user are false.... this user does not actually partake in the mycological community.. all pictures are fabricated pixel by pixel in Adobe Photoshop. This user also accesses the internet from sources other than his own by means of wireless connection. TY

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Re: First AGAR atempt log... [Re: MushroomJoe]
    #5702307 -

Here are the last shots before i go to bed tonight.

Just everything in my glove box and a pic of my new fridge... a little one i picked up for like 60 bucks at the local wally world. great for keeping cultures!!!!



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All posts from this user are false.... this user does not actually partake in the mycological community.. all pictures are fabricated pixel by pixel in Adobe Photoshop. This user also accesses the internet from sources other than his own by means of wireless connection. TY

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Re: First AGAR atempt log... [Re: MushroomJoe]
    #5702470 -

Looks great. That fridge you picked up works perfectly. Looks like it was made for cultures. Keep us posted!


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Hex45 78 70 6c 6f 72 65 20 74 68 65 20 4d 61 67 69
63 2e 20 4b 69 6c 6c 20 79 6f 75 72 20 65 67 6f 21

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Re: First AGAR atempt log... [Re: Kryptik]
    #5702640 -

nice !
jeah i am thinking about getting such a small fridge also,
its really a charm for keeping cultures :smile:

mind your aseptic teks, agar easily contams.


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:penis: MONOTUB tek :sun: HEATBOMB tek :penis:

RIP #cultivation! ....can't associate? well FUCK U !

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Re: First AGAR atempt log... [Re: ohmatic]
    #5702668 -

Did you stuff polyfil into the top of the flask and test tubes? Some sort of filter is required or the agar will draw in contaminants as it shrinks from cooling. Have fun. After you learn agar, you'll wonder how you ever got a crop without it.
RR


--------------------
Download Let's Grow Mushrooms



semper in excretia sumus solim profundum variat

"I've never had a failed experiment.  I've only discovered 10,000 methods which do not work."
Thomas Edison

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Re: First AGAR atempt log... [Re: RogerRabbit]
    #5702923 -

and youll experience the "drowing in agar plates" syndrome lol :smirk:


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:penis: MONOTUB tek :sun: HEATBOMB tek :penis:

RIP #cultivation! ....can't associate? well FUCK U !

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Re: First AGAR atempt log... [Re: ohmatic]
    #5703058 -

The one shelf in the fridge i made out of a piece of Styrofoam that was in the box.. i just cut some holes to hold the test tubed in...

RR... ummmm shit.. lol.. no i did not use poly fill but i did wrap in foil and quickly placed into my glove box after taking out of my PC. so ill just cross my fingers... definitely good info to know though.... THANKS

MJ


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All posts from this user are false.... this user does not actually partake in the mycological community.. all pictures are fabricated pixel by pixel in Adobe Photoshop. This user also accesses the internet from sources other than his own by means of wireless connection. TY

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Re: First AGAR atempt log... [Re: MushroomJoe]
    #5703363 -

I PC agar in a used liquor bottle. Find one with a good screw on lid, and drill a small hole in the lid. You can either stuff it with polyfil or cut a synthetic filter disk to fit inside the small lid. It needs to be filtered or it will contaminate. Be sure to wear surgical gloves that have been washed with alcohol when pouring the plates.
RR


--------------------
Download Let's Grow Mushrooms



semper in excretia sumus solim profundum variat

"I've never had a failed experiment.  I've only discovered 10,000 methods which do not work."
Thomas Edison

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Re: First AGAR atempt log... [Re: MushroomJoe]
    #5703675 -

good luck! I'm in the process of attempting an isolation of one strain (so far, not bad...no contams). Once my greenhouse is up and running, I'll be working with cloning store bought oysters, and when I figure out straw/wood...shiitake cloning will happen.


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channel your inner Larry David

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Re: First AGAR atempt log... [Re: demiu5]
    #5704478 -

Thanks for all the tips RR.. I'm sure I will be coming to you for much advise... (all greatly appreciated)

Well, as soon as i get home today i will be using a multi spore syringes to nocc up these dishes and get them rolling.. my plan right now is to inoc like 2 or 3 per type (meaning one set of HAWAIIAN and one of PF). Would it be necessary to place them in ziploc bags together and then into the incubator or would they be just fine with para-film alone in the incubator??? Thanks again...

MJ


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All posts from this user are false.... this user does not actually partake in the mycological community.. all pictures are fabricated pixel by pixel in Adobe Photoshop. This user also accesses the internet from sources other than his own by means of wireless connection. TY

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Re: First AGAR atempt log... [Re: MushroomJoe]
    #5704581 -

room temp works great. they'll be fine with parafilm. baggies not needed, at least not for incubation...haven't gotten to preservation yet.


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channel your inner Larry David

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Re: First AGAR atempt log... [Re: demiu5]
    #5756222 -

Well, I have had some serious attacks of bacteria. so to counter that effect i have made a second batch with an antibiotic agent mixed in.. will post a new thread for this soon... but i managed to cut and transfer some of the areas and am getting some good results from the transfer.... here are some pics of the transfer.. once i get some more progress on the growth of the transfer i will post pics also..



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All posts from this user are false.... this user does not actually partake in the mycological community.. all pictures are fabricated pixel by pixel in Adobe Photoshop. This user also accesses the internet from sources other than his own by means of wireless connection. TY

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Re: First AGAR atempt log... [Re: MushroomJoe]
    #5756262 -

how did you made the first pic in your last post???

nice work!


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Come see our country, but don't stay too long :smile:

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Re: First AGAR atempt log... [Re: RogerRabbit]
    #5756349 -

RogerRabbit said:
I PC agar in a used liquor bottle. Find one with a good screw on lid, and drill a small hole in the lid. You can either stuff it with polyfil or cut a synthetic filter disk to fit inside the small lid.


i use an old extra vergina oil bottle for my agar,
i simply stuff the opening with polyfill, no need for the lid imho.


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:penis: MONOTUB tek :sun: HEATBOMB tek :penis:

RIP #cultivation! ....can't associate? well FUCK U !

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Re: First AGAR atempt log... [Re: ohmatic]
    #5756424 -

i use canning jars for my agar, except i put the center disk on the correct way ( seal down ) the vaccume on opening isnt a problem, as it is in a glove box anyway.


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ISO: Orissa, Malabar, z strain

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Re: First AGAR atempt log... [Re: ChromeCrow]
    #5756459 -

I agree with demius. Keep petri dishes at room temperature. Don't incubate. Parafilm is all you need. I use four squares per dish and fold it in half lengthwise before wrapping, so it's two layers thick. This prevents the sharp edges of the petri dish from tearing it over time.

A good trick for separating healthy mycelium from bacteria is to pour a fresh layer of hot antibiotic agar right over the top of the contaminated dish. Completely cover the bacteria and mycelium with a fresh layer. The hot agar seems to pasteurize the bacteria, which knocks it back for a few days, but doesn't bother the mushroom mycelium too much, which then grows right up through the agar to appear on top in a couple of days. At the first sight of mycelium on top of the dish, take your scalpel and carefully shave a bit of the mycelium off without taking any of the agar itself. Transfer the myc you shaved to a new dish, and let it grow.

If the bacterial contamination was especially bad, such as in your first pic of the last series above, you might want to repeat the procedure one more time just to make sure. Sometimes bacterial rods can attach to the mycelium as it grows and travel with it.
RR


--------------------
Download Let's Grow Mushrooms



semper in excretia sumus solim profundum variat

"I've never had a failed experiment.  I've only discovered 10,000 methods which do not work."
Thomas Edison

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Re: First AGAR atempt log... [Re: RogerRabbit]
    #5756692 -

I took that pic with my teeth... lol nah just messing.. i had my best friend take it for me.

I am moving them to room temp as we speak.. thanks for all of the advice. All very good things to know


--------------------
All posts from this user are false.... this user does not actually partake in the mycological community.. all pictures are fabricated pixel by pixel in Adobe Photoshop. This user also accesses the internet from sources other than his own by means of wireless connection. TY

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Re: First AGAR atempt log... [Re: RogerRabbit]
    #7573167 -

RogerRabbit said:
A good trick for separating healthy mycelium from bacteria is to pour a fresh layer of hot antibiotic agar right over the top of the contaminated dish.  Completely cover the bacteria and mycelium with a fresh layer.  The hot agar seems to pasteurize the bacteria, which knocks it back for a few days, but doesn't bother the mushroom mycelium too much, which then grows right up through the agar to appear on top in a couple of days.  At the first sight of mycelium on top of the dish, take your scalpel and carefully shave a bit of the mycelium off without taking any of the agar itself.  Transfer the myc you shaved to a new dish, and let it grow. 
RR


when you say hot agar do you mean as hot as when you normaly pour dishes? and do I use just agar or the agar plus nutriens mix (MEA, PDA...)?

Many thanks! :grin::thumbup:


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Make your own Magnetic Stirrer for L.C ( Several designes Links)

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Re: First AGAR atempt log... [Re: FGL]
    #7575441 -

:confused:???????????????????


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Make your own Magnetic Stirrer for L.C ( Several designes Links)

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Re: First AGAR atempt log... [Re: FGL]
    #7575499 -

Thats some neat advice. I think your doing just fine sofar


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PF TEK - writeup by EvilMushroom666
Lets Grow Mushrooms - RogerRabbit & RoadKills website with sample videos plus the full PF TEK video series. Alot of great information - BUY THE DVD
Cakes can and will pin! - So you think cakes suck for pins. Your wrong
Franks Simple Coir/Verm Tek
Franks Proper Pasturisation Tek
Franks Spawning To Bulk - Monotub
Professor Pinheads RTV Injection Port Tek
Foo Mans No Soak WBS Prep Tek

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Re: First AGAR atempt log... [Re: veda_sticks]
    #7575517 -

sorry man...have you read my questions? what aam i doing fine so far?:rolleyes:


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Make your own Magnetic Stirrer for L.C ( Several designes Links)

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Re: First AGAR atempt log... [Re: FGL]
    #7575610 -

lol, just commenting that while u have questions and a few problems it looks like your doing a not to bad job so far.

If it where my first attempt, i probably be throwing petri dishes of green mold away.


--------------------

PF TEK - writeup by EvilMushroom666
Lets Grow Mushrooms - RogerRabbit & RoadKills website with sample videos plus the full PF TEK video series. Alot of great information - BUY THE DVD
Cakes can and will pin! - So you think cakes suck for pins. Your wrong
Franks Simple Coir/Verm Tek
Franks Proper Pasturisation Tek
Franks Spawning To Bulk - Monotub
Professor Pinheads RTV Injection Port Tek
Foo Mans No Soak WBS Prep Tek

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Re: First AGAR atempt log... [Re: veda_sticks]
    #7575625 -

FGL
Hot MEA with Gentmycin sulphate as antibiotic is what I use. Normally, I let the agar cool until I can hold the bottle to pour without any heat protection for my hands. If I'm trying to pour over bacterial contamination, I'll pour it a bit hotter. The idea is to knock the bacteria back with heat, but the stronger mushroom mycelium will survive the hot agar to grow up through to the top.
RR


--------------------
Download Let's Grow Mushrooms



semper in excretia sumus solim profundum variat

"I've never had a failed experiment.  I've only discovered 10,000 methods which do not work."
Thomas Edison

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Re: First AGAR atempt log... [Re: veda_sticks]
    #7575648 -

veda_sticks said:
lol, just commenting that while u have questions and a few problems it looks like your doing a not to bad job so far.

If it where my first attempt, i probably be throwing petri dishes of green mold away.


Ahhh:grin: but this is not my first attempt but MushroomJoe (the original poster) first attempt...:tongue::grin:


--------------------
Make your own Magnetic Stirrer for L.C ( Several designes Links)

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Re: First AGAR atempt log... [Re: RogerRabbit]
    #7575652 -

RogerRabbit said:
FGL
Hot MEA with Gentmycin sulphate as antibiotic is what I use.  Normally, I let the agar cool until I can hold the bottle to pour without any heat protection for my hands. If I'm trying to pour over bacterial contamination, I'll pour it a bit hotter.  The idea is to knock the bacteria back with heat, but the stronger mushroom mycelium will survive the hot agar to grow up through to the top.
RR


Perfect!:thumbup:
Many thanks for the answer RR!;):thumbup::thumbup::thumbup:


--------------------
Make your own Magnetic Stirrer for L.C ( Several designes Links)

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