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OfflineChode420
Stranger
Registered: 12/08/05
Posts: 6
Last seen: 17 years, 6 months
Just ziplock bags and grain invitro
    #5035922 - 12/08/05 08:11 PM (18 years, 3 months ago)

unfortunately i cant experiment right now cause i'm on my holiday in thailand :P...anyway i'm having an idea of a Very easy,alternative way without pressure cooking and using only big,thick ziplock bags and grain(i'm thinking to use rye).I used several teks from Mycotopia for this idea.
so here's the plan:
1.soak grain for 24h(endospores will start to germinate), simmer until its dehydrated enough and let it drain for a while.
2.fill ziplock bag half way or 1/3 with grain
3.use fractional sterilisation :
---------------------------------------------------------
1) Steam heating to 100 ?C for 30 min
Vegetative cells are destroyed but endospores survive
2) Incubate at 30?C-37?C overnight
Most bacterial endospores germinate
3) Second heat treatment, 100 ?C, 30 min
Germinated spores are killed
4) Second incubation at 30?C-37 ?C overnight
Remaining endospores germinate
5) Third heat treatment, 100 ?C, 60 min
Last remaining germinated endospores are killed
--------------------------------------------------
4.let bags cool and take 'em to glovebox, then use a paper piercer to make some holes to the bag and cover them with 'band aid tape'
5. clean a spot on the bag and innoc. then cover with tape.
6.let the bags incubate 'til 100%
7.then do G2G to a bigger ziplock or just use the same colonized bags: brake the substrate into nice chunks, fill the bag with clean water and dunk for the night.
8.next day use several dunked bags, cut a hole in the bottom, drain and put the chunks in to a another, bigger ziplock.this time dont make any filter holes, just open the siplock only VERY little bit for the air exchange.
and maybe put a small layer of coco coir or whatever casing material on top.
9.let incubate for few days and maybe...get a reasonable crop!


let me know what do you think.can the ziplock bag even take the boiling temp or will it melt?is this method even worth to try???

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OfflineThai_connection
Siam
 User Gallery

Registered: 10/01/05
Posts: 247
Loc: Thailand
Last seen: 13 years, 5 months
Re: Just ziplock bags and grain invitro [Re: Chode420]
    #5036785 - 12/08/05 10:59 PM (18 years, 3 months ago)

don't worry...if you going to the south island you can ask for cubies with your omeltes and other food....just ask for "hedd mou"
peace
siam

Edited by Thai_connection (12/08/05 11:00 PM)

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Invisiblemicro
bunbun has a gungun
Male User Gallery

Registered: 05/09/03
Posts: 7,532
Loc: Brick City Flag
Re: Just ziplock bags and grain invitro [Re: Chode420]
    #5037833 - 12/09/05 04:38 AM (18 years, 3 months ago)

Is it possible it could work?

Sure....

Is it a hell of lot more likely to fail?

Yeah....

If you have sterile mycelium water it's a lot more likely to work because you could inject a lot into it and not have to wait for germination, mix it up and it will colonise faster, but still it's a bit chinchy....

If you ARE going to do it that way the goal is to colonise it as fast as you can while still being sterile. Mixing it up at least two times would be really benificial (once at first and once at like 15-25% colonised); just be really careful not to introduce contaminants that way. Even if you had a couple spores invade, unless it's actually pathogenic, competitors could be overrun by the "good" mycelium.


--------------------
Any research paper or book for free
(Avatar is Maxxy, a character by Mizzyam, RIP)

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OfflineChode420
Stranger
Registered: 12/08/05
Posts: 6
Last seen: 17 years, 6 months
Re: Just ziplock bags and grain invitro [Re: Chode420]
    #5038504 - 12/09/05 09:27 AM (18 years, 3 months ago)

I'm thinking now to fill only 1/3 of the bag (a small bag with a very small amount of grain)and let air inside and close it.. so there will be no need for the air holes and then inoc. with liquid culture.the small amount of grain will colonize very fast without the need for new air. make lots of these bags and later mix in to a bigger one.
this should keep it more clean and even more easyer.  :grin:

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Invisibleagar
old hand
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Registered: 11/21/04
Posts: 9,056
Loc: Somewhere Else
Trusted Cultivator
Re: Just ziplock bags and grain invitro [Re: Chode420]
    #5038870 - 12/09/05 10:47 AM (18 years, 3 months ago)

You are in the WRONG FORUM.

Learn a few things about what you might propose.

Then, you might not propose them.

As, you would know, they are impractical.


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

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OfflineChode420
Stranger
Registered: 12/08/05
Posts: 6
Last seen: 17 years, 6 months
Re: Just ziplock bags and grain invitro [Re: agar]
    #5041287 - 12/09/05 08:44 PM (18 years, 3 months ago)

maybe i am in the wrong forum coz u guys experiment how to make bigger flushes etc. Actually i HAVE learned a few things and grown using different methods.
my point is not to have a greenhouse or growchamber...just basic materials.
i know that if you do a proper casing its gonna be nice flush, but it's gonna be stealth and invitro for me this time.i have been reading a lot and found proof for my ideas(you really CAN use ziplock bags..and you really CAN grow myc without air holes if there is enough air inside and if its gonna colonize fast enough and etc..... )
im not looking for very big flushes from my bags.

but maybe i am an idiot..then please correct me when i'm tottally wrong, i just wanna see if i have contams or not.... :smile:

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