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Invisiblejellyfish

Registered: 10/02/05
Posts: 7,457
Grain to Grain
    #6558687 - 02/12/07 03:15 PM (17 years, 1 month ago)

My friend inoculated 8 popcorn jars that had a polyfill filter and a innoc hole covered with micropore tape under a flow hood after spraying lysol and took many other sterile precautions. Only one jar germinated. It is colonizing fast but took 3 weeks to get going. Discouraged he decided to do some pf tek cakes with coffee and made a shitty glovebox using a tupperware container and ducktape (he figured that as long as there was no air flow into the container he good keep in fairly sterile with his lysol. All of the pf tek jars got contams. He is out of spores (strain is b+) so his only hope is using the mycellium on his nearly colonized popcorn jar.
He has tried doing LCs but he has got contams everytime. Could it be because of unsterile micropore tape? Is there anyway to use polyfill with an LC? He figures he should crumbe up the popcorn cake and put a piece in some rye jars but he is afraid of contams. Is there anyway for him tp get the mycellium into the rye jar without a glovebox? He really doesn't want to lose his myc but agar has always failed him.

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Invisibleodium33
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Registered: 05/16/06
Posts: 1,187
Loc: mycoland
Re: Grain to Grain [Re: jellyfish]
    #6558711 - 02/12/07 03:20 PM (17 years, 1 month ago)

prepare a quart jar half full with sterile water, and a tyvek filter, in a a cheap glove box, dump water into colonized jar, shake it up and dump it back into more sterile water. that gives you enough lc to inoculate hundreds of jars. then with the rest of the colonized jar, grain to grain 1:10 to new jars of sterile substrate


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Invisiblejellyfish

Registered: 10/02/05
Posts: 7,457
Re: Grain to Grain [Re: odium33]
    #6558759 - 02/12/07 03:35 PM (17 years, 1 month ago)

Since the last glove box failed miserably what materials should the new one be made from? I also don't know where to find tyvek in Canada thats why i use polyfill

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Invisibleodium33
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Registered: 05/16/06
Posts: 1,187
Loc: mycoland
Re: Grain to Grain [Re: jellyfish]
    #6558769 - 02/12/07 03:38 PM (17 years, 1 month ago)




6 dollar box from walmart, piece of plexiglass, some trash bag scraps and a polyfilled hole. i just recently did a 120 jar G2G with this glove box with 0% contamination


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Invisiblejellyfish

Registered: 10/02/05
Posts: 7,457
Re: Grain to Grain [Re: odium33]
    #6558822 - 02/12/07 03:58 PM (17 years, 1 month ago)

What kind of gloves do you have and how are they attached? My friends ws pretty ackward to use. How do you sterilze it? How do silicone injection ports work can I use them instead of micropore tape and how safe are they against contams. Do I just put the silicone on once or after I innoc too?

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Invisibleodium33
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Registered: 05/16/06
Posts: 1,187
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Re: Grain to Grain [Re: jellyfish]
    #6559754 - 02/12/07 06:47 PM (17 years, 1 month ago)

i just wear some purple nitrile gloves and put my arms under the trash bags hanging down. a glovebox just has to stop the airflow in the room. i find attached gloves to be way more trouble then they are worth. i have never worked with silicone ports but my understanding is that you prep your lid with 2 holes, fill one with polyfil, or cover with some sort of filter material (tyvek) then u put a blob of silicone over the other hole and inject through that, and it reheals itself.


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Invisiblejellyfish

Registered: 10/02/05
Posts: 7,457
Re: Grain to Grain [Re: odium33]
    #6559933 - 02/12/07 07:26 PM (17 years, 1 month ago)

K thanks a lot

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Offlineshroomsynergy
Fungi Freak
Male

Registered: 01/17/06
Posts: 861
Loc: Andromeda
Last seen: 4 years, 6 months
Re: Grain to Grain [Re: jellyfish]
    #6560112 - 02/12/07 08:15 PM (17 years, 1 month ago)

most postal offices have free tyvek envelopes. that sucks that you guys are out of spores. alot of people here have B+, and i'm sure that one of them would be willing to hook you up with a print or syringe.

you may want to check out the oven tek, or just wait to get some printable caps from that jars yeild.


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InvisibleBlutjager
Inhuman
Male

Registered: 06/11/06
Posts: 9,220
Re: Grain to Grain [Re: shroomsynergy]
    #6560407 - 02/12/07 09:14 PM (17 years, 1 month ago)

You don't have to go crazy building a glove box,some of my most successful inoculations ever have taken place in an upside down glass aquarium.Put everything you will be using in it and spray the hell out of everything with a 10-12% bleach solution,no gloves no nothing,remember its still air not sterile air in the glovebox

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Invisiblejellyfish

Registered: 10/02/05
Posts: 7,457
Re: Grain to Grain [Re: Blutjager]
    #6571705 - 02/15/07 05:37 PM (17 years, 1 month ago)

Thing is the glove box was fairly well sealed and the gloves were completly attached. And there was a 100% failure 7 pf tek jars and 1 lc. He's never been able to do an lc and it's been tried many times. I don't know why the hell it's failing so badly he has done one succesful popcorn jar out of 6 and have done pf teks before that were 100% successful. There is no way he will be able to save the mycellium from his jar when he sucks this bad at growing but we can't pinpint the problems. Flowhood, mask, gloves, alcohol soaked cottonballs for the syringe,lysol on all the surfaces etc etc.
I read about people here who aren't too anal about sterilization and get good results but he can't even get agar colonized. It starts to colonize and then bam! contams. I'll tell him to try the oven tek and to make an LC from the jar itself and use the popcorn to inncoulate some rye jars but I feel that as soon as the lid of the jar cracks the rye will be contamed. I was thinking maybe he should just grow the popcorn out and get a print but theres no way he'll be able to do it contamless.

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Invisibleagar
old hand
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Registered: 11/21/04
Posts: 9,056
Loc: Somewhere Else
Trusted Cultivator
Re: Grain to Grain [Re: Blutjager]
    #6571762 - 02/15/07 05:56 PM (17 years, 1 month ago)

Quote:

upside down glass aquarium



How do you get your arms in an upside-down aquarium?
(assuming its upside-down on a solid surface)


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InvisibleBlutjager
Inhuman
Male

Registered: 06/11/06
Posts: 9,220
Re: Grain to Grain [Re: agar]
    #6571814 - 02/15/07 06:11 PM (17 years, 1 month ago)

Quote:

agar said:
Quote:

upside down glass aquarium



How do you get your arms in an upside-down aquarium?
(assuming its upside-down on a solid surface)






Just pull it over the edge of whatever its sitting on,put everything your going to use in it,spray everything with a bleach solution and I'm good to go :rockon:



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Invisibleodium33
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Registered: 05/16/06
Posts: 1,187
Loc: mycoland
Re: Grain to Grain [Re: Blutjager]
    #6571946 - 02/15/07 06:47 PM (17 years, 1 month ago)

Quote:

Blutjager said:
Quote:

agar said:
Quote:

upside down glass aquarium



How do you get your arms in an upside-down aquarium?
(assuming its upside-down on a solid surface)






Just pull it over the edge of whatever its sitting on,put everything your going to use in it,spray everything with a bleach solution and I'm good to go :rockon:







damn lol thats a good idea.  my first aquarium glove box was turned on its side, with a trash bag cover, with another layer underneath it with arm holes.  it was terribly ghetto but it worked lol


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OfflineBigSistah
Stranger
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Registered: 10/17/06
Posts: 38
Last seen: 17 years, 1 month
Re: Grain to Grain [Re: jellyfish]
    #6572215 - 02/15/07 07:51 PM (17 years, 1 month ago)

As one LC and grain failure to another – they’re both hard! I do everything in an oven that’s been heated to 400 and cooled down, and I’ve managed to get two good LCs and one good grain jar in a year. Great luck spawning from cakes, though, and enough spore prints to last a lifetime.

Anyway, being experienced in salvaging disasters, I’d suggest that you diversify with what you do with your remaining asset. In addition to trying the previously suggested leachate, get that grain cased and fruiting, and transfer a couple of nice chunks of myc into small PF jars. Sometimes it works, sometimes it doesn’t, but outcomes are better if you let the chunks swim in a hydrogen peroxide solution (1:5) for a minute prior to poking them in with sterile forceps. Once you have fruits, you can clone (hard) or make a spore syringe (easy) to save your strain.

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