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OfflineNeuronicObserver
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Reinoculate if jars do not colonize?
    #16182794 - 05/04/12 06:06 PM (12 years, 8 months ago)

1. I have a few jars which have not began colonization. It seems like a good idea to just inoculate them again to save resources/hassle. Does this seem valid? It has been a couple of weeks, so either some of the spores are very slow at germinating or they are not viable or something else went wrong. There seems to be stark inconsistency with these MS spores, it is disconcerting :/

Other questions that I have not found answers to:
2. Has anyone read about or tried using hybrid grade verm? I mean mixing fine verm with medium/coarser verm. I thought of this for giving the fine verm more structural integrity against the compaction caused by pressure cooking. Also this would theoretically give the substrate best of both worlds. Some of my jars have this, waiting to measure results against control group.
3. Is compaction from pressure cooking a widespread issue and what kind of solutions have you guys come up with? I have read that you should pack the jars more tightly since they will compact upon cooking anyways, which prevents open space above the dry verm layer. I am using an 8 quart presto pressure cooker.


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OfflineMr. Wilson
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Re: Reinoculate if jars do not colonize? [Re: NeuronicObserver]
    #16182806 - 05/04/12 06:08 PM (12 years, 8 months ago)

this your first go round??


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OfflineNeuronicObserver
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Re: Reinoculate if jars do not colonize? [Re: Mr. Wilson]
    #16182822 - 05/04/12 06:12 PM (12 years, 8 months ago)

Quote:

Mr. Wilson said:
this your first go round??




2nd. The previous grow was sterilized by boiling a pot of water. I have now moved onto a pressure cooker and it has given me some trouble.


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Edited by NeuronicObserver (05/04/12 06:12 PM)

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OfflineMr. Wilson
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Re: Reinoculate if jars do not colonize? [Re: NeuronicObserver]
    #16182827 - 05/04/12 06:13 PM (12 years, 8 months ago)

was the first a success??


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OfflineNeuronicObserver
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Re: Reinoculate if jars do not colonize? [Re: Mr. Wilson]
    #16182839 - 05/04/12 06:16 PM (12 years, 8 months ago)

Quote:

Mr. Wilson said:
was the first a success??




Yes, but I also had slowish/inconsistent colonization even though I used the correct substrate ratios, temperature, and 1-2ml of spore solution per jar.


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Edited by NeuronicObserver (05/04/12 06:20 PM)

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OfflineMr. Wilson
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Re: Reinoculate if jars do not colonize? [Re: NeuronicObserver]
    #16182882 - 05/04/12 06:24 PM (12 years, 8 months ago)

how long did you PC the jars at 15 POUNDS--how long did they cool---is the foil or some other obstruction still on top of jars--when did you inoculate??? questions questions---usually 15 people done jumped on this trying to help i feel im being watched--ill get you straight though


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OfflineSimpleFarmer
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Re: Reinoculate if jars do not colonize? [Re: Mr. Wilson]
    #16182894 - 05/04/12 06:26 PM (12 years, 8 months ago)

Your gettin evaluated  :smile:

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OfflineMr. Wilson
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Re: Reinoculate if jars do not colonize? [Re: SimpleFarmer]
    #16182910 - 05/04/12 06:30 PM (12 years, 8 months ago)

i fuckin KNEW it---yall boys are always fast round here


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OfflineNeuronicObserver
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Re: Reinoculate if jars do not colonize? [Re: Mr. Wilson]
    #16182914 - 05/04/12 06:31 PM (12 years, 8 months ago)

Quote:

Mr. Wilson said:
how long did you PC the jars at 15 POUNDS--how long did they cool---is the foil or some other obstruction still on top of jars--when did you inoculate??? questions questions---usually 15 people done jumped on this trying to help i feel im being watched--ill get you straight though




This makes me very happy, thanks for offering your insight :smile:
The jars were PC'd for 60 minutes, PSI is not indicated, so I'm assuming it is 15 PSI. They cooled for about 2 days, then I inoculated them. I might be PCing 10 more jars in case there is something wrong with my current batch. Do you think there is anything I should do different with these?


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OfflineMr. Wilson
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Re: Reinoculate if jars do not colonize? [Re: NeuronicObserver]
    #16182928 - 05/04/12 06:36 PM (12 years, 8 months ago)

is the foil still on em---two weeks is more than enough time for contams to start---i assume the jars were sterile---i have never had spores take that long to pop though but its not unheard of--keep em at room temp on a shelf for another week or so---you really need to get a PC with a gauge on it though---id bet you left the foil on though


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Edited by Mr. Wilson (05/04/12 06:41 PM)

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OfflineNeuronicObserver
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Re: Reinoculate if jars do not colonize? [Re: Mr. Wilson]
    #16182937 - 05/04/12 06:40 PM (12 years, 8 months ago)

Quote:

Mr. Wilson said:
is the foil still on em




Nope, there is just masking tape covering the holes.


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OfflineMr. Wilson
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Re: Reinoculate if jars do not colonize? [Re: NeuronicObserver]
    #16182948 - 05/04/12 06:43 PM (12 years, 8 months ago)

THERES ya problem---take that shit off---i put a coffee filter on mine then screw the band down and leave it till the spores pop---then take it off---worked real good for me----them spores HAVE to have some air man---if theres any contams in there they're gonna pop too though---but its unavoidable---save ya syringe---if thangs dont work out i might still be able to help you


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Edited by Mr. Wilson (05/04/12 06:48 PM)

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OfflineNeuronicObserver
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Re: Reinoculate if jars do not colonize? [Re: Mr. Wilson]
    #16182995 - 05/04/12 06:55 PM (12 years, 8 months ago)

Quote:

Mr. Wilson said:
THERES ya problem---take that shit off---i put a coffee filter on mine then screw the band down and leave it till the spores pop---then take it off---worked real good for me----them spores HAVE to have some air man---if theres any contams in there they're gonna pop too though---but its unavoidable---save ya syringe---if thangs dont work out i might still be able to help you




I would like to try that, but I fear that taking off the lids to install the coffee filters right now would invite contams. I could still take the tape off though right?


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OfflineMr. Wilson
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Re: Reinoculate if jars do not colonize? [Re: NeuronicObserver]
    #16183013 - 05/04/12 06:58 PM (12 years, 8 months ago)

you put the lids on seal side down didnt you--dont matter nyway thats just something i do--but yeah man take that tape off and give em a week--something will SURELY happen


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Edited by Mr. Wilson (05/04/12 07:10 PM)

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OfflineNeuronicObserver
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Re: Reinoculate if jars do not colonize? [Re: Mr. Wilson]
    #16183084 - 05/04/12 07:16 PM (12 years, 8 months ago)

Quote:

Mr. Wilson said:
you put the lids on seal side down didnt you--dont matter nyway thats just something i do--but yeah man take that tape off and give em a week--something will SURELY happen




I did put them on seal side down. Is there really be that large of a difference? I'll detape them. And out of curiosity, is it ever a good idea to reinoculate?


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OfflineMr. Wilson
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Re: Reinoculate if jars do not colonize? [Re: NeuronicObserver]
    #16183419 - 05/04/12 08:39 PM (12 years, 8 months ago)

seal side down means that lid is pretty much glued to the glass---youll know what i mean when you birth em---no worries--i had a bit of a fungus gnat problem thats how i learned bout the coffee filters---dont reinnoc though--just do what i said--if you reshoot em youll proly have TOO much water--and i got my doubts on your pc method nyway---trust me just take the tape off-put em up-and leave em alone for a week--TRUST ME---this is the batch i have now---shot 11--birthed 11--no glove box---put in the FC last niteif you got spores galore i would just start another batch


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Edited by Mr. Wilson (05/04/12 08:45 PM)

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InvisibleThayendanegea
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Re: Reinoculate if jars do not colonize? [Re: Mr. Wilson]
    #16183454 - 05/04/12 08:50 PM (12 years, 8 months ago)

Go to corner drug store and buy some micro pore tape...cost you like 2 bucks....remove the masking tape and replace it with that....do not take lids off.
As far as re-innoc....I had a jar that was filled with substrate and pc'd but ran out of ms so it sat for 3 months....knocked it up with next batch and did great.


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OfflineMr. Wilson
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Re: Reinoculate if jars do not colonize? [Re: Thayendanegea]
    #16183508 - 05/04/12 09:01 PM (12 years, 8 months ago)

micro dont breathe as good as some think it does---that verm layer is really sufficient in itself---but anykind of tape or anything is gonna hinder gas exchange--even the coffee filter to some extent


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OfflineThe Influence
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Re: Reinoculate if jars do not colonize? [Re: Mr. Wilson]
    #16183851 - 05/04/12 10:15 PM (12 years, 8 months ago)

Your PC method sounds fine, obviously those jars are sterile otherwise you would have contams growing.

Did you shake your syringe between each jar? Either way those jars should have shown germination. I would toss them and start over. Never re-inoculate, you have no idea how much water has evaporated or hasn't so shooting them up could throw moisture content off. BRF verm is cheap so just start over.

Also tape is really only used to stop water from getting in during sterilization. I used to use micropore tape and leave it on. It does not stop GE, as my jars are fully colonized between 2-3 weeks and that has been consistent even after removing the tape. Coffee filters will not stop contams whatsoever so using them is futile and opening your jar to put them on is just un-needed risk.Verm is a great contaminant barrier, I have done dozens of grows now and never a contam once.

If your pc has a weight make sure both weights are on and that will be 15psi. What make of PC do you got?

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Re: Reinoculate if jars do not colonize? [Re: The Influence]
    #16183883 - 05/04/12 10:23 PM (12 years, 8 months ago)

im curious too why its been two weeks and no action--ive heard of slow spores -just not seen em yet--he should have seen something by now--slow spores+no GE=double whammy??--im still kind of a noob too---id like to see the guy get something for his effort--whether boomers or a learning experience


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Edited by Mr. Wilson (05/04/12 10:30 PM)

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OfflineThe Influence
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Re: Reinoculate if jars do not colonize? [Re: Mr. Wilson] * 1
    #16184161 - 05/04/12 11:25 PM (12 years, 8 months ago)

Quote:

Mr. Wilson said:
im curious too why its been two weeks and no action--ive heard of slow spores -just not seen em yet--he should have seen something by now--slow spores+no GE=double whammy??--im still kind of a noob too---id like to see the guy get something for his effort--whether boomers or a learning experience



Yeah I'm scratching my head as well. I am no expert but have been at this awhile now and never had this happen. Personally though I would just throw them out, maybe take the tape off and wait a few more days but to me it's already been to long lol, I just freaked cause the jars I got colonizing now took 6 days to show signs of growth. I have gotten used to seeing growth in my jars from an ms inoculation in 3-4 days. It sounds like some of his jars are going so it's not a total loss.

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Invisiblecubes4cancer
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Re: Reinoculate if jars do not colonize? [Re: The Influence]
    #16184204 - 05/04/12 11:35 PM (12 years, 8 months ago)

i think op innoced the dry verm layer in these jars,if you didnt watch each innoc point to see if the spore water ran down the side of the glass,its probably the case.its happened to me before,i just reinnoced soon after i reckonized my problem. :thumbup:


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ROGERRABBIT: P cubensis will grow on stuffed animals, bibles, quarter pounds of weed, bras, etc.
In other words,it's hard to fuck up a cube grow.
RR

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Re: Reinoculate if jars do not colonize? [Re: cubes4cancer]
    #16184504 - 05/05/12 12:59 AM (12 years, 8 months ago)

Quote:

cubes4cancer said:
i think op innoced the dry verm layer in these jars,if you didnt watch each innoc point to see if the spore water ran down the side of the glass,its probably the case.its happened to me before,i just reinnoced soon after i reckonized my problem. :thumbup:



Hhhm maybe he did, but in that case that is even riskier business to re-inoculate then. Your dry verm layer is well supposed to be dry so that any contaminate spores can not germinate and colonize down to the substrate. If this is the case I would definitely say throw them out and that you sir got very lucky.

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OfflineMr. Wilson
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Re: Reinoculate if jars do not colonize? [Re: NeuronicObserver]
    #16185094 - 05/05/12 07:31 AM (12 years, 8 months ago)

well neuronicO what have you done with em?? inquiring minds want to know--


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Re: Reinoculate if jars do not colonize? [Re: Mr. Wilson]
    #16185133 - 05/05/12 07:46 AM (12 years, 8 months ago)

i had the same problem awhile back 99% of mine was lack of air exchange took regular tape off and put on micro pore and everything started mycing up

also mabey lack of correct temp?

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Re: Reinoculate if jars do not colonize? [Re: The Influence]
    #16185158 - 05/05/12 07:56 AM (12 years, 8 months ago)

how would you solve the temp issue if that was it??  aargh--meant for bulsie


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Edited by Mr. Wilson (05/05/12 07:57 AM)

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OfflineRogerRabbitM
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Re: Reinoculate if jars do not colonize? [Re: Mr. Wilson]
    #16185284 - 05/05/12 09:08 AM (12 years, 8 months ago)

Quote:

Mr. Wilson said:
micro dont breathe as good as some think it does---that verm layer is really sufficient in itself---but anykind of tape or anything is gonna hinder gas exchange--even the coffee filter to some extent




Nonsense.

Gasses will easily penetrate micropore tape.  Masking tape is a different story.  Coffee filters in mycology are dumb-They're like trying to catch mosquitoes with a butterfly net.

To the OP, two points to consider:
1.  Did you allow steam to vent for several minutes before sealing up the pressure cooker to allow it to build pressure?  If you failed to do this, you had 15 psi of air pressure and it won't do squat.  You need 15 psi of steam pressure to sterilize and the only way to get this is to allow all the air to vent out first before sealing the unit.

2. Never re-inoculate.  If the jars didn't colonize, there's a reason.  Make up new jars and try again.
RR


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Re: Reinoculate if jars do not colonize? [Re: RogerRabbit]
    #16185369 - 05/05/12 09:47 AM (12 years, 8 months ago)

well there you go OP--thats the final word--RR-i use the coffee filters to catch dem bastard fungus gnats--and it just makes me feel better--and yeah you gotta let that steam start coming out the tube good and heavy before you put the jiggler on--thats in the instructions of your PC OP--wish ya the best of success--and i would solve a colonizing temp issue by just leaving em on a shelf at room temp--i still believe it was the masking tape that gotcha--good luck


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Edited by Mr. Wilson (05/05/12 09:54 AM)

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OfflineNeuronicObserver
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Re: Reinoculate if jars do not colonize? [Re: RogerRabbit]
    #16185393 - 05/05/12 09:54 AM (12 years, 8 months ago)

Temperature is right at 80F

I'll be getting some micropore tape today.

I am reasonably sure that the inoculation went past the dry verm layer, I'll watch that more closely with this next PC batch.

Using this PC: http://www.gopresto.com/products/products.php?stock=01282

@RR, the design of this PC seems to manage the steam build up automatically. I followed the directions for it, there was no indication of needing to allow steam buildup prior to sealing.

Notes:
A few of the jars that are having trouble were compacted from PC, so there is a gap between the dry verm layer and lid. This was made worse by how the dry verm is fine grade, it fell down the sides of the jars to some extent. This however doesn't solve the problem because there are some new jars which have a much smaller gap and no exterior verm fallout.

With my third PC, I'll make sure the jars are extra compact(within reason), to eliminate the gap 100%. I'm not sure whether the verm will fall down again if I use fine, so maybe I should stick with coarse for the verm layer?

Thanks guys for all of the advice, I feel welcomed :smile:


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OfflineNeuronicObserver
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Re: Reinoculate if jars do not colonize? [Re: Mr. Wilson]
    #16185408 - 05/05/12 09:58 AM (12 years, 8 months ago)

Quote:

Mr. Wilson said:
well there you go OP--thats the final word--RR-i use the coffee filters to catch dem bastard fungus gnats--and it just makes me feel better--and yeah you gotta let that steam start coming out the tube good and heavy before you put the jiggler on--thats in the instructions of your PC OP--wish ya the best of success--and i would solve a colonizing temp issue by just leaving em on a shelf at room temp--i still believe it was the masking tape that gotcha--good luck




I'm looking at the manual right now. It says to seal the pressure cooker and put on the pressure regulator(jiggler) prior to heating.


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OfflineMr. Wilson
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Re: Reinoculate if jars do not colonize? [Re: NeuronicObserver]
    #16185437 - 05/05/12 10:09 AM (12 years, 8 months ago)

hmmm--i just wait till that steam starts comin out and grab the jiggler with a dish towel and put it on--wait till you hit 15 lbs-reduce the heat a little--like from high to bout med high--start timer-i run for 75 minutes--i have NO problems


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OfflineNeuronicObserver
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Re: Reinoculate if jars do not colonize? [Re: Mr. Wilson]
    #16185471 - 05/05/12 10:21 AM (12 years, 8 months ago)

Quote:

Mr. Wilson said:
hmmm--i just wait till that steam starts comin out and grab the jiggler with a dish towel and put it on--wait till you hit 15 lbs-reduce the heat a little--like from high to bout med high--start timer-i run for 75 minutes--i have NO problems




I'll try it :smile:


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Re: Reinoculate if jars do not colonize? [Re: NeuronicObserver]
    #16185491 - 05/05/12 10:25 AM (12 years, 8 months ago)

got time--wanna start another run--ill babysit ya and make sure you get it right--im sitting here thinking bout starting a run myself


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OfflineNeuronicObserver
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Re: Reinoculate if jars do not colonize? [Re: Mr. Wilson]
    #16185686 - 05/05/12 11:20 AM (12 years, 8 months ago)

Quote:

Mr. Wilson said:
got time--wanna start another run--ill babysit ya and make sure you get it right--im sitting here thinking bout starting a run myself




I just have to fight through this final exam for college, then I'll be free to mycologize! I'm hoping to PC at around 6 or 7 today. Btw your jars are looking beautiful.


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Re: Reinoculate if jars do not colonize? [Re: Mr. Wilson]
    #16185694 - 05/05/12 11:23 AM (12 years, 8 months ago)

Pure fucking nonsense  !!! Lol

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Re: Reinoculate if jars do not colonize? [Re: SimpleFarmer]
    #16186047 - 05/05/12 01:03 PM (12 years, 8 months ago)

whaaa???


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Re: Reinoculate if jars do not colonize? [Re: Mr. Wilson]
    #16186060 - 05/05/12 01:06 PM (12 years, 8 months ago)

Lol just giving ya shit

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Re: Reinoculate if jars do not colonize? [Re: SimpleFarmer]
    #16186094 - 05/05/12 01:13 PM (12 years, 8 months ago)

tell me what you really think about me directing this dude-please dont be politr--im thick skinned-i can deal--i just wanna help as much as yall helped me--if im an idiot and just got lucky this time just tell me--lol


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Re: Reinoculate if jars do not colonize? [Re: Mr. Wilson]
    #16186104 - 05/05/12 01:16 PM (12 years, 8 months ago)

Quote:

Mr. Wilson said:
hmmm--i just wait till that steam starts comin out and grab the jiggler with a dish towel and put it on--wait till you hit 15 lbs-reduce the heat a little--like from high to bout med high--start timer-i run for 75 minutes--i have NO problems




This is the correct method, even if the directions for the pressure cooker say to put the weight on first.
RR


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Download Let's Grow Mushrooms



semper in excretia sumus solim profundum variat

"I've never had a failed experiment.  I've only discovered 10,000 methods which do not work."
Thomas Edison

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OfflineSimpleFarmer
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Re: Reinoculate if jars do not colonize? [Re: Mr. Wilson]
    #16186122 - 05/05/12 01:21 PM (12 years, 8 months ago)

Your doing fine man , I just like giving advice on things I know and can show them a trusted cultivator  said so too .. I know people bitch about people that don't use the search bar but I like it bc I can go and search for what they are asking for and hook them up with the link and in the process learn it myself :wink: a lot of people call it spoon feeding but I call it helpful research ..you are doing good man keep up the good work

Edited by SimpleFarmer (05/05/12 01:25 PM)

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OfflineMr. Wilson
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Re: Reinoculate if jars do not colonize? [Re: SimpleFarmer]
    #16186166 - 05/05/12 01:33 PM (12 years, 8 months ago)

cool RR-i know if i dont know any thing bout shroom growing--i KNOW im an expert at running a PC--me and my folks grow a HELL of a garden every year---and ill get disowned if i buy ANY kind of vegtable out of a grocery store that can be grown here


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OfflineMr. Wilson
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Re: Reinoculate if jars do not colonize? [Re: SimpleFarmer]
    #16186224 - 05/05/12 01:48 PM (12 years, 8 months ago)

i believe i got my colonizing game down--its that fruiting game im lacking in--alot of people have a tendency to overcomplicate things myself included--this thing really is simple---provided you do it the RIGHT way---heard a sayin once----when your're young goin through life hammering down -thinking your blazing a new trail one day you get older then slow down you get to look around -you look down and see somebody else's footprints


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OfflineSimpleFarmer
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Re: Reinoculate if jars do not colonize? [Re: Mr. Wilson]
    #16186260 - 05/05/12 02:02 PM (12 years, 8 months ago)

Nice :thumbup:

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OfflineMr. Wilson
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Re: Reinoculate if jars do not colonize? [Re: SimpleFarmer]
    #16186265 - 05/05/12 02:04 PM (12 years, 8 months ago)

i learned a while back when the wise man is tellin you how to do something you do it the way he says---hence why i woulndt know what little i do know if it werent for you guys---this place is a fuckin wellspring


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Edited by Mr. Wilson (05/05/12 02:06 PM)

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OfflineXeonen
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Re: Reinoculate if jars do not colonize? [Re: Mr. Wilson]
    #16186385 - 05/05/12 02:41 PM (12 years, 8 months ago)

Quote:

Mr. Wilson said:
im curious too why its been two weeks and no action--ive heard of slow spores -just not seen em yet--he should have seen something by now--slow spores+no GE=double whammy??--im still kind of a noob too---id like to see the guy get something for his effort--whether boomers or a learning experience





Once, I've inoculated 8 jars with P. Ostrateus, in those days I had a gf interested in growing mushrooms so I gave her a jar. After two weeks of waiting I've seen no growth in jars and tossed them.

Interestingly enough, after two weeks (total of one month) after inoculation my ex told me she started to see "white spots" growing within the jar.

Well, sometimes they take it real SLOW.


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OfflineMr. Wilson
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Re: Reinoculate if jars do not colonize? [Re: Xeonen]
    #16187363 - 05/05/12 06:42 PM (12 years, 8 months ago)

ayy yay yay--in my lifes experiences those thangs that take the longest are usually the best


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OfflineThe Influence
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Re: Reinoculate if jars do not colonize? [Re: Mr. Wilson]
    #16187416 - 05/05/12 06:56 PM (12 years, 8 months ago)

Quote:

Mr. Wilson said:
ayy yay yay--in my lifes experiences those thangs that take the longest are usually the best



You've never had sex while drunk I take it :lol:

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OfflineMr. Wilson
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Re: Reinoculate if jars do not colonize? [Re: The Influence]
    #16187467 - 05/05/12 07:12 PM (12 years, 8 months ago)

allright peckerneck--lol--hey check my new post in cultivation lets chat awhile


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OfflineMr. Wilson
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Re: Reinoculate if jars do not colonize? [Re: Mr. Wilson]
    #16187499 - 05/05/12 07:21 PM (12 years, 8 months ago)

hey NeuroticO where you at--lets get this party started--hey i might make some grain jars if these fellas talk me into it


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