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Amazon Shop for: ½ Pint Jars, 10cc Syringe, Brown Rice Flour, Vermiculite

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InvisibleSam1912
journeyman

Registered: 09/23/04
Posts: 2,142
Loc: Cali
GT on speed trial
    #3238048 - 10/10/04 02:41 AM (12 years, 2 months ago)

Here's an experiment to minimize the colonization time. I'd love to see it duplicated on a more aggressive strain to comparison purposes.

Purpose: Achieve minimum colonization time using Golden Teacher Strain.

Goal: Aquire at least 1 fully colonized jar before time specified.

Conditions of Failure:
--1. More than 1 contaminated jar.
--2. Time to full colonization greater than 8 days 3 hours.
--3. Divergence from ideal colonization conditions.

Ideal conditions to be maintained:
--1. Presence of Dextrose in spore syringe.
--2. Presence of Dextrose in substrate
--3. Constant incubation temperatue of 84F
--4. High CO2 level during the entire incubation period.

Materials:
--1. Sterile water(5% dextrose,.9% NaCl)
--2. BRF
--3. Vermiculite
--4. 10cc Spore syringe
--5. mineral water
--6. sterile equipment(i.e. gloves, scalpels, container, etc.)
--7. CO2 cartridges
--8. 1/2 Pint jars with 9 holes on each (4 outside, 4 staggered on inside, 1 in the middle)
--9. Aluminum foil

Procedures:

<Jars>
--1. PF jars were prepared in 270cc jars using 1/4 cup BRF, 1/2 cup Vermiculite, 30cc of mineral water. Top vermiculite layer was ground to very find grade.
--2. Jars were sterilized using steam for 3 hours with tapes and aluminum foil.
--3. Sterilized jars were incubated for 2 days to screen possible contamination.
<spore syringes>
--4. 10cc syringe(dark and no dextrose) was dispensed into a sterile container and was diluted to 40cc.
--5. New spore solution was loaded into sterile 10cc syringes and was allowed to germinate for 24 hours and time matched to end of monitoring the jars.
<CO2>
--6. A sterile container was filled with CO2.
--7. 4 - 10cc syringes were filled with gas from the container.(I couldn't put gas directly into the syringes without possibly compromising the sterility.)
<inoculation>
--8. Tapes and foil were removed from the jar.
--9. Using 1.25" needle, 1 CO2 syringe was dispensed into the jar through the middle hole.
--10. 1 syringe with 1.5" needle containing water(5%dextrose) was dispensed as evenly as possible throughout the substrate.
--11. 1 syringe containing prepared spore solution was dispensed 1cc per site using outer and inner holes and 2cc through the center hole.
--12. Aluminum foil was replaced and allowed for beginning of colonization.

<colonization>
--after each inspection, bleed some CO2 into the incubator. Stop CO2 supplement at 20% colonization from outside.

--------------------------
It's been 7 and a half hours now since beginning of colonization and about 31 hours since I started touching these spores. I plan on giving some fresh air at around 25% and 75% by flipping the jars for 5 min. Idea being tested here is dex(which I'm pretty sure helps germination) and high CO2 at early stage. I'll post updates every time I check.

Sam


--------------------
Protect your civil rights!  End drug prohibition.  And if you don't care about your civil rights, protect mine!

If you want a rating from me, please PM me.  For those really newbies, don't expect an answer back, but you can try me anyways.


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Offlineaussieone
shroominator
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Registered: 07/11/04
Posts: 766
Loc: QLD, Australia
Last seen: 2 years, 2 months
Re: GT on speed trial [Re: Sam1912]
    #3238264 - 10/10/04 04:18 AM (12 years, 2 months ago)

the fastest ive got my jars to colonize is 12days GT on 1lt millet, temp at level 28deg, using a liquid culture would cut the time down considerably


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


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Offlinescatmanrav
Brainy Smurf

Registered: 05/08/04
Posts: 11,483
Loc: Flag
Last seen: 3 years, 10 months
Trusted Cultivator
Re: GT on speed trial [Re: aussieone]
    #3238270 - 10/10/04 04:22 AM (12 years, 2 months ago)

Just use a liquid culture. Just cased some GT's done 8 days (actually 7 1/2) after injection. 3cc's and your golden :thumbup:

And co2 won't do anything, the myc already produces enough within its environment easily for what it needs.


--------------------
"life is like a drop of rain getting closer and closer to falling into a lake, and then when you hit the lake there is no more rain drop, only the lake."

Growing with bags, start to finish (including my new grain and substrate prep)
Anyone looking to start bulk tubs/mono tubs/shotgun hybrids? Good tubs to use..
How I do grain (old still good tips)
Turn your closet into a fruiting chamber
Casing layer colonization and overlay


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Offlineaussieone
shroominator
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Registered: 07/11/04
Posts: 766
Loc: QLD, Australia
Last seen: 2 years, 2 months
Re: GT on speed trial [Re: scatmanrav]
    #3238276 - 10/10/04 04:31 AM (12 years, 2 months ago)

^^^ yep if anything the extra water content might breed contams


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


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InvisibleSam1912
journeyman

Registered: 09/23/04
Posts: 2,142
Loc: Cali
Re: GT on speed trial [Re: aussieone]
    #3238553 - 10/10/04 07:43 AM (12 years, 2 months ago)

It's an experiment. There's only ambient CO2 in the jar when you inoculate and the small mycelium after couple of days shouldn't have produced a lot of CO2. Air should still be rather fresh at that point. Trying to see if I can kick start with CO2. If so, I can try CO2 in other places, too.

As far as water content... max fruiting formula is 60 cc of water. I used 30cc while mixing, 10cc dex solution, and another 10 cc spores. That's 50cc. It looked pretty good. Didn't really look dry after only 50cc.

Two environmental factors for the colonization are being tested to see if it make a difference. 8days 3 hours is how long it took to get 2 of my 15 GT jars to get done before.

Anyways, thanx for at least reading.

Sam


--------------------
Protect your civil rights!  End drug prohibition.  And if you don't care about your civil rights, protect mine!

If you want a rating from me, please PM me.  For those really newbies, don't expect an answer back, but you can try me anyways.


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InvisibleSam1912
journeyman

Registered: 09/23/04
Posts: 2,142
Loc: Cali
Re: GT on speed trial [Re: Sam1912]
    #3250815 - 10/13/04 04:50 AM (12 years, 1 month ago)

you know what's bad about innoculating on 9 sites? Whole damn thing germinates. It's patchy white everywhere. It looks like "mostly cloudy sky". Only thing that I can imagine using to gauge progress in the bottom.

I can't update... The whole damn jar germinated. I'll dissect a jar in few days and give you guys an update.

Sam


--------------------
Protect your civil rights!  End drug prohibition.  And if you don't care about your civil rights, protect mine!

If you want a rating from me, please PM me.  For those really newbies, don't expect an answer back, but you can try me anyways.


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Offlinescatmanrav
Brainy Smurf

Registered: 05/08/04
Posts: 11,483
Loc: Flag
Last seen: 3 years, 10 months
Trusted Cultivator
Re: GT on speed trial [Re: Sam1912]
    #3250870 - 10/13/04 05:32 AM (12 years, 1 month ago)

Germinated? I'm confused..patchy white everywhere? Mycelium germinated? Isn't that good?

Yeah I'm confused.


--------------------
"life is like a drop of rain getting closer and closer to falling into a lake, and then when you hit the lake there is no more rain drop, only the lake."

Growing with bags, start to finish (including my new grain and substrate prep)
Anyone looking to start bulk tubs/mono tubs/shotgun hybrids? Good tubs to use..
How I do grain (old still good tips)
Turn your closet into a fruiting chamber
Casing layer colonization and overlay


Post Extras: Print Post  Remind Me! Notify Moderator
InvisibleSam1912
journeyman

Registered: 09/23/04
Posts: 2,142
Loc: Cali
Re: GT on speed trial [Re: scatmanrav]
    #3250879 - 10/13/04 05:36 AM (12 years, 1 month ago)

damn.. no I meant spores germinated then grew fuzz. I feel like being picked and poked on. Damn it! I'm already frustrated with this experiment as is... Maybe I'll just throw the stupid jars out.

I'm sure you would tell me that, too, right, scatmanrav?

Sam


--------------------
Protect your civil rights!  End drug prohibition.  And if you don't care about your civil rights, protect mine!

If you want a rating from me, please PM me.  For those really newbies, don't expect an answer back, but you can try me anyways.


Post Extras: Print Post  Remind Me! Notify Moderator
Offlinescatmanrav
Brainy Smurf

Registered: 05/08/04
Posts: 11,483
Loc: Flag
Last seen: 3 years, 10 months
Trusted Cultivator
Re: GT on speed trial [Re: Sam1912]
    #3250911 - 10/13/04 05:51 AM (12 years, 1 month ago)

If spores germinated and grew fuzz wouldnt that be myc? Or did you mean they germinated and started growing myc then fuzzed?

Or are you talking about the syringe you made and havnt knocked up the jars yet? If the spores germinated in the solution, thats fine..even better actually..

So I guess I'm still confused?


--------------------
"life is like a drop of rain getting closer and closer to falling into a lake, and then when you hit the lake there is no more rain drop, only the lake."

Growing with bags, start to finish (including my new grain and substrate prep)
Anyone looking to start bulk tubs/mono tubs/shotgun hybrids? Good tubs to use..
How I do grain (old still good tips)
Turn your closet into a fruiting chamber
Casing layer colonization and overlay


Post Extras: Print Post  Remind Me! Notify Moderator
InvisibleSam1912
journeyman

Registered: 09/23/04
Posts: 2,142
Loc: Cali
Re: GT on speed trial [Re: scatmanrav]
    #3250935 - 10/13/04 06:02 AM (12 years, 1 month ago)

okay.. I spore syringe I inoculated with didn't have any visible sign of germination. But I saw fuzz everywhere on the glass surface(sides, not bottom). I didn't really check on these until few hours ago. So, I'm grouping fuzz with germination. probably poor choice of word to describe what's going on, but it's my first peek(no, really, first peek) and it's really frustrating to know that I'll have no clue when it'll be considered 100% colonized.

I probably should've thought about how I'm going to gauge progress before I put 10cc(2cc from original syringe i made +8cc water with dex) of spore solution in the jar. I'm probably going to just leave it in the incubator until I dissect them. It was just an experiment... but i think I met the condition of failure because I won't be able to tell their progress. Don't even know 25% and 75% to when i was supposed to flip the jars...

So, there. This was just plain stupid experiment.


--------------------
Protect your civil rights!  End drug prohibition.  And if you don't care about your civil rights, protect mine!

If you want a rating from me, please PM me.  For those really newbies, don't expect an answer back, but you can try me anyways.


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Offlineoddiseus
The PsychofunkFirefly

Registered: 10/05/04
Posts: 456
Loc: Doldrums & Faultlines
Last seen: 3 years, 1 month
Re: GT on speed trial [Re: Sam1912]
    #3252331 - 10/19/04 06:25 PM (12 years, 1 month ago)

I think that ultimately if your jars went 100% before the expected time of 8 days 3 hrs(in your case) than you actually proved that it works. Don't give up yet see it through then fruit em !!!! its my HO
=)


--------------------
Though you can't go back and make a new beginning. You can start today to make a new end.

Traveller, there is no path, paths are made by walking

If Jesus was jewish...Why does he have a mexican name??


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Amazon Shop for: ½ Pint Jars, 10cc Syringe, Brown Rice Flour, Vermiculite

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