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Invisibleoontribe
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inoculating grain jars
    #21774429 - 06/07/15 02:39 PM (8 years, 7 months ago)

Two days ago i inoculated 14 quart millet jars with only 6ccs of MS syringe.
some were inoculated with half cc or less!
yea i know few drops has thousands of spores but i think that iam still under the recommended amount off ccs/jar.
do i need to reinoculate em? Or Iam good.
And i gave em a shock right after the inoculation process.


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Offlinelivelogikal
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Re: inoculating grain jars [Re: oontribe]
    #21774458 - 06/07/15 02:45 PM (8 years, 7 months ago)

Here's the thing brotha! You might throw off your moisture with that amount! typically 1-2ml is all you need! More than that you are adding to much water back to your grain! Definitely do not add more and next time use much less!!


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Offlinetetherface
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Re: inoculating grain jars [Re: livelogikal]
    #21774897 - 06/07/15 04:19 PM (8 years, 7 months ago)

1cc is the same as 1ml  i always use around 1cc/ml per jar but have gotten away with much less im sure you'll be fine


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Invisibleoontribe
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Re: inoculating grain jars [Re: tetherface]
    #21776893 - 06/08/15 12:45 AM (8 years, 7 months ago)

Thanks guys but is it going to affect anything?


Edited by oontribe (06/08/15 01:06 AM)


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OfflineMachiavelliavore
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Re: inoculating grain jars [Re: oontribe]
    #21777016 - 06/08/15 01:39 AM (8 years, 7 months ago)

You should be fine.  My understanding is that generally you don't want to shake after spore inoculation, as the spores germinate by connecting to other spores, thus you want them all in a few cluster.

I think most germination occurs from spore flakes though, so it probably won't end up mattering.


After you see some decent size bright white streaks (say, 20% of the volume colonized,) shake the jars again thoroughly.  The idea is to have enough colonized grains to effectively inoculate the whole jar.  If you don't, stalls are quite likely.  It may take a while hit that 20%~ mark with such a small ammount of spore solution, personally I'd accept the few stalls you're likely to get rather than trying a second delayed innocculation.

Also, throwing off the moistre content of a quart jar with half a cc of water?  That's silly.  Even 2cc's wouldn't matter.


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I spawned some popcorn casings and had double-overlay cause I didn't put enough hydrogen peroxide in my automated aquarium mister.  I only got one mushroom so I cut off the head part where the seeds fall from and put it in a jar of LC and sprayed it all over a tin of PF cakes I made with gravel, cardboard, and bisquick in my microwave.  I think it will be good cause B+ is so potent.
Triggered yet?

Only a square would say "a cube is a cube."


No, this does not look right...


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Invisibleoontribe
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Re: inoculating grain jars [Re: Machiavelliavore]
    #21777195 - 06/08/15 03:24 AM (8 years, 7 months ago)

So slower colonization...thanks man.


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Offlineblackout
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Re: inoculating grain jars [Re: livelogikal]
    #21777235 - 06/08/15 03:55 AM (8 years, 7 months ago)

Quote:

livelogikal said:
Here's the thing brotha! You might throw off your moisture with that amount! typically 1-2ml is all you need! More than that you are adding to much water back to your grain! Definitely do not add more and next time use much less!!



He used 6cc over 14 jars, not 6cc per jar.


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OfflineMachiavelliavore
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Re: inoculating grain jars [Re: oontribe]
    #21777242 - 06/08/15 04:00 AM (8 years, 7 months ago)

Probably depends on spore concentration in the syringe and the aggresiveness of the strain, but all other factors being equal, I would expect the immediate shake + small quantity of spore solution to slow down the time to reach that critical 15-20% for the shake.  After the shake it should be about the same.

If you inoculated through silicon injection ports or pfill with a clean syringe and flame sterilized, even if they are slow they probably won't contaminate or stall.  If you popped the lids or used a syringe from a homemade print, it's more likely to contaminate (esp bacteria.)

Beating contamination is based mostly on statistical advantage and/or cleanliness.  Spore solution is fairly poor statistically, especially in such a large jar, but luckily it can be done cleanly with relative ease.

I suppose I should ammend and say that if it's a homemade syringe from a non-invitro open-air print, a second round of double-tap innoculations might be worthwhile

I'm not a pro, but I'm fairly certain this information is correct (from my experience and reading.)


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I spawned some popcorn casings and had double-overlay cause I didn't put enough hydrogen peroxide in my automated aquarium mister.  I only got one mushroom so I cut off the head part where the seeds fall from and put it in a jar of LC and sprayed it all over a tin of PF cakes I made with gravel, cardboard, and bisquick in my microwave.  I think it will be good cause B+ is so potent.
Triggered yet?

Only a square would say "a cube is a cube."


No, this does not look right...


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Invisibleoontribe
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Re: inoculating grain jars [Re: Machiavelliavore]
    #21777274 - 06/08/15 04:20 AM (8 years, 7 months ago)

The syringe was fairly loaded with spores as the water looked purple its made out of vendor's print.
before inoculating the jars i bombed the room and the SAB with oust alchol and bleach, took a shower brushed my teeth, clean clothes, facemask, rubber gloves.
filter is poly stuffed in 1/4" hole, but i opened the jars lid to inoculate.
am i in risk to lose the jars?
and if i am going to re-inoculate throw the poly filter do i need to use a SAB?
and i ll have to flame sterilise the needle after each jar?


Edited by oontribe (06/08/15 05:05 AM)


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OfflineMachiavelliavore
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Re: inoculating grain jars [Re: oontribe]
    #21777447 - 06/08/15 06:02 AM (8 years, 7 months ago)

Quote:

oontribe said:
The syringe was fairly loaded with spores as the water looked purple its made out of vendor's print.
before inoculating the jars i bombed the room and the SAB with oust alchol and bleach, took a shower brushed my teeth, clean clothes, facemask, rubber gloves.
filter is poly stuffed in 1/4" hole, but i opened the jars lid to inoculate.
am i in risk to lose the jars?
It sounds like you were very careful and used SAB, so the lid pop will probably not result in contamination.  Since you shook, any contaminant will be spead out, and re-innoculating probably won't help.
and if i am going to re-inoculate throw the poly filter do i need to use a SAB?
No, but it's advisable to use the SAB anyway.  I haven't used poly, but you probably do need to wait a few seconds to avoid melting it with the hot needle.  I don't know why you didn't noc through the poly, as that is the main benefit of poly over tyvek.
and i ll have to flame sterilise the needle after each jar?
No reason not to other than laziness that I'm aware of.





I think you should be fine with your current inoculation.  There's borderline no risk of adding a contaminant that isn't already in your spawn jars using a vendor syringe through poly in an SAB, unless the vendor syringe is contaminated.  If your initial lid pops did introduce a contaminant, it's unlikely that a delayed second spore inoculation will overtake it, and IMO it would be preferable to see that contaminant germinate and save the rest of your spore syringe.

If you have the time in the future, nocing two half-full quart jars then G2G'ing might be preferable to such a massive spore inoculation.  Getting a clean print without an invitro grow may be tricky, and your clean vendor syringe will last many grows if you G2G.

P.S.  It will probably take longer than typical to SEE your spore germination, since the shake spread out your spores.  Many of them will be in the interior where you cannot see, and they will also be less dense, thus harder to see at the beginning.  That doesn't mean it will be slower, just a different development pattern of a little bit everywhere instead of a lot in two spots.  One squirt down opposite walls is the typical inoculation pattern.  Once you hit 15-20% you shake and effectively G2G the rest of the jar.


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


I spawned some popcorn casings and had double-overlay cause I didn't put enough hydrogen peroxide in my automated aquarium mister.  I only got one mushroom so I cut off the head part where the seeds fall from and put it in a jar of LC and sprayed it all over a tin of PF cakes I made with gravel, cardboard, and bisquick in my microwave.  I think it will be good cause B+ is so potent.
Triggered yet?

Only a square would say "a cube is a cube."


No, this does not look right...


Edited by Machiavelliavore (06/08/15 06:12 AM)


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Invisibleoontribe
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Re: inoculating grain jars [Re: Machiavelliavore]
    #21777975 - 06/08/15 09:42 AM (8 years, 7 months ago)

Thanks for the information my friend.


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Re: inoculating grain jars [Re: oontribe]
    #21791544 - 06/11/15 02:28 AM (8 years, 7 months ago)

So 6 days after inoculating 14 jars with only 6cc and shaking them after inoculation.
js this going good or?


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OfflineShroominMe
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Re: inoculating grain jars [Re: oontribe]
    #21791746 - 06/11/15 04:46 AM (8 years, 7 months ago)

Not good.  Great!  :headbang:


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Re: inoculating grain jars [Re: ShroominMe]
    #21791790 - 06/11/15 05:22 AM (8 years, 7 months ago)

Lool thx man


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Re: inoculating grain jars [Re: oontribe]
    #21792012 - 06/11/15 07:24 AM (8 years, 7 months ago)

In the future for best results don't shake right after inoculating with spores.

LC-shake
LI-shake
Agar wedges-shake
spore solution-no shake

all of the above - give a good shake when jars are around 20% or so colonized to distribute mycelium....:thumbup:


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Edited by FreeWorldOrder (06/11/15 07:24 AM)


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Invisibleoontribe
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Re: inoculating grain jars [Re: FreeWorldOrder]
    #21792080 - 06/11/15 08:04 AM (8 years, 7 months ago)

awsome...
i have a question.
what would happen if i shake before the 20% mark


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OfflineMachiavelliavore
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Re: inoculating grain jars [Re: oontribe]
    #21793458 - 06/11/15 03:19 PM (8 years, 7 months ago)

You will shock and break up the forming mycellium, but there will not be enough to be effectively spread around the whole jar, meaning your colonization will be slow or you will need to shake a second time later.
Since each shake has a 1-3 day recovery period just for the mycellium to get rhizo/fluffy and start colonizing new grain again, you really want to wait until you have enough that a shake will result in having a colonized grain nearby (say penny radius) of any non-colonized grain.


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


I spawned some popcorn casings and had double-overlay cause I didn't put enough hydrogen peroxide in my automated aquarium mister.  I only got one mushroom so I cut off the head part where the seeds fall from and put it in a jar of LC and sprayed it all over a tin of PF cakes I made with gravel, cardboard, and bisquick in my microwave.  I think it will be good cause B+ is so potent.
Triggered yet?

Only a square would say "a cube is a cube."


No, this does not look right...


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Invisibleoontribe
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Re: inoculating grain jars [Re: Machiavelliavore]
    #21793498 - 06/11/15 03:28 PM (8 years, 7 months ago)

Thanks machia


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