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Offlinebmarley3434
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bit off more than i can chew AHHH
    #3423156 - 11/29/04 03:36 PM (12 years, 14 days ago)

well i just recieved 3 cultures.. oysters reishi and shiitake.. in hopes that i would be confident enough by the time they got here. and to force me to just TRY it(ive been reading for months). but that couldnt be farther from the truth.. im determined not to let these cultures contam or i will feel like such a bastard. this is my first time so bare with me if you can  :confused:..

1. how do i transfer from culture to substrate? only methods i can find on this is for psilocybes which require jars. i am pretty sure i can skip this method and inoculate a substrate bag correct?

2. what is an efficient way of maintaining proper temperatures? would a temperature controlled space heater work? or would that fluctuate too much . and is it ok to run one of those things all the time

well these are my main questions for now and thanks all of you for your insight and patience


--------------------
www.returntonature.us - wild foods blog


OM


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Offlineragadinks
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Re: bit off more than i can chew AHHH [Re: bmarley3434]
    #3423303 - 11/29/04 04:11 PM (12 years, 14 days ago)

Quote:


1. how do i transfer from culture to substrate? only methods i can find on this is for psilocybes which require jars. i am pretty sure i can skip this method and inoculate a substrate bag correct?



Have you got agar cultures ? If yes, you only have to transfer them from the petri dish to the sterilized substrate bag under a laminar flow or in a glovebox. Then seal the bag so that no contams can enter.

Quote:


2. what is an efficient way of maintaining proper temperatures? would a temperature controlled space heater work? or would that fluctuate too much . and is it ok to run one of those things all the time



When you have sealed your mycobags well you only have to put them in a warm room where the temp is about 24-26 degrees Celsius.


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Offlinebmarley3434
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Re: bit off more than i can chew AHHH [Re: ragadinks]
    #3424306 - 11/29/04 07:57 PM (12 years, 14 days ago)

how long will the ajar dishes last till it contams if in the fridge..

so with temp when it talks about what induces pinning and what induces fruiting.. is that just optimal.. will everything go fine at a consistent temp of 24-26

http://www.mycobags.com/frameset.html << reliable vendor?

will the same substrate work fine for oysters to shittake to reishi..

i really appreciate it raga i cant thank you enough


--------------------
www.returntonature.us - wild foods blog


OM


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InvisiblePeterthinks
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Registered: 11/10/04
Posts: 2,378
Re: bit off more than i can chew AHHH [Re: bmarley3434]
    #3424811 - 11/29/04 09:31 PM (12 years, 14 days ago)

don't forget to save a little bit in some slants so you can do it again.
And don't use the very outside of the dish...that's usually where the contaminants are hiding if they're there...they won't show till they get in the bag if they're hiding.
here are the growing conditions you need to set up for your mushrooms.
http://www.sacredshrooms.org/forums/showflat.php/Cat/0/Number/2103080/an/0/page/0#2103080
the oyster may fruit at a constant temp but don't count on it.
I'd make more cultures out of what I got before I'd fruit.You can lose everything in the blink of an eye.
here is a link of the basics.
http://www.mykoweb.com/articles/cultivation.html
Just mellow and don't open the cultures till you have something to propogate it into.
and if you use grain soak it in warm water overnight to wake up all the bacteria...THEN pressure cook it,If you just add water then pressure cook it you'll keep getting mystery contaminations!
:jester:
Don't vacuum the day before,shut off the furnace/AC around 4 am and do your transfers in a little used room(put towels over the vents too)first thing in the morning before the dust gets stirred up.
the night before clean the work area with %5 bleach/water solution and have everything ready for the morning.
You should be able to walk in,wipe down again...sit down...and do your transfers without any messing around.Wear clean clothes,have your hair covered and don't breathe onto your cultures.
You can work without a flow hood or glovebox...you just have to be really REALLY anal about it.
Good luck:jester:


Edited by Peterthinks (11/29/04 09:48 PM)


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Offlineragadinks
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Re: bit off more than i can chew AHHH [Re: Peterthinks]
    #3426107 - 11/30/04 02:14 AM (12 years, 13 days ago)

Yeah, Peterthinks has said all you can say ;-)
Wish you luck and post some pics if you have success ...


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Offlinebmarley3434
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Re: bit off more than i can chew AHHH [Re: ragadinks]
    #3432544 - 12/01/04 02:02 PM (12 years, 12 days ago)

"Have you got agar cultures ? If yes, you only have to transfer them from the petri dish to the sterilized substrate bag under a laminar flow or in a glovebox. Then seal the bag so that no contams can enter."


does anyone have a tek on this?


--------------------
www.returntonature.us - wild foods blog


OM


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Offlinebmarley3434
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Re: bit off more than i can chew AHHH [Re: bmarley3434]
    #3434480 - 12/01/04 09:32 PM (12 years, 12 days ago)

can i ac plastic petri dishes? or will they melt


--------------------
www.returntonature.us - wild foods blog


OM


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Offlineragadinks
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Re: bit off more than i can chew AHHH [Re: bmarley3434]
    #3436523 - 12/02/04 11:08 AM (12 years, 11 days ago)

> does anyone have a tek on this?
I do not know of any, but I am sure you will find some stuff if you search with google or on the shroomery ...

> can i ac plastic petri dishes? or will they melt
No you cannot ac them - they will melt !


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Offlinejmeuser
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Registered: 11/13/04
Posts: 32
Loc: Golden, Colorado
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Re: bit off more than i can chew AHHH [Re: ragadinks]
    #3439078 - 12/02/04 08:09 PM (12 years, 11 days ago)

1. I have always gone to spawn jars and then substrate. It is always good to have multiple points of inoculation and this is accomplished by either rye or millet grains. Ace brand bird seed works great because it is mostly millet. Just pour boiling water over it, let it soak over night, and then sterilize (I use an autoclave but a pressure canner is the preferred at-home method) You'll need filters for the jars but it is the same with psylocybe. After your jars are colonized then go to spawn bags. Until your ready just throw the plates in the fridge, they'll be fine. Also, I've developed a at-home tech for making sterile plates using a microwave that I can share with you.

2. The absolutely chillest way of keeping your cultures warm is so simple and beautiful. I have a gas oven with a pilot light. The pilot light keeps the oven at about 24 degrees C, even though my house is a chilly 5 degrees, brrrrr. My oven fits about six 5 pound mushroom bags. Another tech is to put an aquarium heater in a large bin, then put your fungus in another container within this bin. I use this method for my spawn jars.

Good luck and feel to PM with me with any specific questions.


--------------------
-Jon

Have:
Pleurotus pulmonarius (Pheonix Oyster)
Letinula edodes (Shitake)
Ganoderma lucidum (Reishi)

Want for trade:
Other edibles . . .


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Offlinefalcon
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Re: bit off more than i can chew AHHH [Re: bmarley3434]
    #3439327 - 12/02/04 08:58 PM (12 years, 11 days ago)

Practice how you will make the
transfers before you do it.

You want to make smooth steady movements when you
make the transfer. It really helps if you go over
how you will hold the scalpel, how you will
flame the scalpel, in what order
you will do what, ect.

Most contmination falls onto the agar if
you are not working under a flow hood
so minimize the time the agar is exposed.
If you make smooth steady motions you
cut down on turbulence near your work.

Wear latex gloves or better yet nitrile.
Practice with the gloves on. Working
with gloves on feels different.
When you clean your hands with isopropal
it drys your skin and creates flakes
that can fall into your work.
With gloves this is less of a problem.

Start with the oyster culture or
the reishi. If you make a mistake
with either of these it is less
likely to fail than the shiitake.

Give it a couple days rest and
do the next culture.
This will give you time to think
about what you did right and
how you can improve your technique.


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Offlineragadinks
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Re: bit off more than i can chew AHHH [Re: jmeuser]
    #3440963 - 12/03/04 05:09 AM (12 years, 10 days ago)

> Also, I've developed a at-home tech for making sterile plates using a microwave that I can share with you.
Yeah, please share it. I am eager to hear about new/different teks ...


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Offlinejmeuser
Eco-logic
Registered: 11/13/04
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Loc: Golden, Colorado
Last seen: 11 years, 5 months
Re: bit off more than i can chew AHHH [Re: ragadinks]
    #3443463 - 12/03/04 08:58 PM (12 years, 10 days ago)

About the microwave tech . ..

I developed a microwave tech at UC Davis while I was a teaching assistant in a plant tissue culture class. The professor I worked for provided me with a somewhat old paper on microwave sterilization (forgive me I have no idea where the paper is now).

The primary trouble with microwaving media is that it quickly boils over even at the lowest setting of the microwave(mw). While trying to use my mw to sterilize my media I ran into this problem many times before I figured out the key. All you have to do is put a few other large containers of water in the microwave with your media. These other containers will soak up much of the microwaves, decreasing the effective wattage on your media. So set your microwave on the lowest setting, put in a few 1L containers of water and microwave the sh*t out of your media (between 10-40 minutes). There are some other minor considerations. 1. You should try it first with water and measure your water loss so you can adjust your protocol accordingly 2. The media may still boil over; you need to watch it, stop it if it boils over, wait a bit, and start microwaving again (ideally the temp. should hover right below boiling for this tech to be optimized and it is far differenct for every microwave, volume of excess water, and container. 3. Your media will probably be too hot to pour when done microwaving, you'll still need a quasi sterile place for your media to cool as cooling will draw air into your container (if you seal your container following microwaving you risk implosion and air will be sucked in when you open it anyway). If you're going 100% ghetto you can still do a good job. Just blast your oven full temp. Turn it off. Then duct tape it at the seams while it cools. When your bottles are done microwaving, spray with ethanol, stick in oven and allow to cool. 4. Lastly, as always never microwave a sealed container or metal (duh, I know). I think that is about it, enjoy it, this is the first time I've shared it.


--------------------
-Jon

Have:
Pleurotus pulmonarius (Pheonix Oyster)
Letinula edodes (Shitake)
Ganoderma lucidum (Reishi)

Want for trade:
Other edibles . . .


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Offlinefalcon
In the green

Registered: 04/01/02
Posts: 6,835
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Re: bit off more than i can chew AHHH [Re: jmeuser]
    #3443826 - 12/03/04 11:12 PM (12 years, 9 days ago)

:thumbup:


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Offlineragadinks
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Re: bit off more than i can chew AHHH [Re: jmeuser]
    #3445214 - 12/04/04 11:12 AM (12 years, 9 days ago)

Thanks for sharing.
That whole procedure sounds reasonable and I will try it the next time I am going to make some agar.
:yesnod:


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Amazon Shop for: ½ Pint Jars, Agar, Aquarium Heater, Autoclave, Laminar Flow Hood, Mycobags, Petri Dish, Rye Grain, Wild Bird Seed

Mushrooms, Mycology and Psychedelics >> Gourmet and Medicinal Mushrooms

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