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OfflineFluzzball
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Inoculation Question
    #21994434 - 07/25/15 09:48 AM (8 years, 6 months ago)

Now, I am a complete noobie and this is my first time ever growing mushrooms, so I am bound to make mistakes for forgive me for my noobnishness!

I was wondering if it is possible to "reinoculate" jars. For instance I am worried that perhaps I inoculated too soon after sterilizing which could have killed the spores. The jars were warm to the touch but were not hot, so I am not sure. If the spores died what will happen inside the jar? Will another culture take over? Or, after about 10 - 15 days if nothing is still happening with mycelium growth and there is no contaminants could I reinoculate or would I have to start from scratch again?

It's only day 2 so I am not expecting much to go down, but I am merely curious if that is a possibility.

I appreciate any information you could provide for me!
Thanks in advance!


Edited by Fluzzball (07/25/15 09:59 AM)


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OfflinePsilosoulful

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Re: Inoculation Question [Re: Fluzzball]
    #21994552 - 07/25/15 10:17 AM (8 years, 6 months ago)

Wait a week, it is probable that some spores died, but many probably survived the heat. Did you inject the MS syringe into grains or brf? A brf jar would have more inoculation points for spores to germinate, that is if you used 4 holes on the lid.


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OfflineFluzzball
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Re: Inoculation Question [Re: Psilosoulful]
    #21994595 - 07/25/15 10:32 AM (8 years, 6 months ago)

I used 4 inoculation points and they are brf cakes. Hopefully some survived!


--------------------
“Everybody is a genius. But if you judge a fish by its ability to climb a tree, it will live its whole life believing that it is stupid.”
- Albert Einstein





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OfflinePsilosoulful

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Re: Inoculation Question [Re: Fluzzball]
    #21994620 - 07/25/15 10:37 AM (8 years, 6 months ago)

Only time will tell. :goodluck:


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OfflineGanjaburger
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Re: Inoculation Question [Re: Psilosoulful]
    #21994646 - 07/25/15 10:46 AM (8 years, 6 months ago)

I've had similar issues myself.... But honestly whether u r using grain, or brf, they r both so cheap. Why not just start fresh? Unless u just knocked up a couple hundred jars then I'd say start fresh to avoid any possible issues later on.  Plus u will gain experience everytime u have to "re-do" something.

    Look at it as knowledge gained.  In this hobby u can never have to much knowledge!  Good luck, and the only other tip I can give u is...... Patience is crucial!

Ganjaburger


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


Trade /sell list...plenty of cube prints....microscope, two hundred dollar brand new Aerobed, and a ti-84+ calculator....


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OfflinePsilosoulful

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Re: Inoculation Question [Re: Ganjaburger]
    #21994661 - 07/25/15 10:51 AM (8 years, 6 months ago)

:whathesaid: You should have multiple jars going at the same time, so if you lose one it isn't a big deal. And next time wait for the jars to cool down completely!! It's not worth the stress of mixing brf and PC'ing it, to mess it up at the inoculation stage.


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OfflineFluzzball
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Re: Inoculation Question [Re: Psilosoulful]
    #22000107 - 07/26/15 02:02 PM (8 years, 6 months ago)

Thanks everyone for your help! I will wait it out and see how things turn out. I inoculated 10 jars with the verm/brown rice flour method. Day 3 and still nothing. I am going to put them somewhere warmer, as they are only at about 70 degrees and I read keeping them at about 75-85 is an ideal temperature. Figured I'd find a bin, put some water in a bowl, use a fish tank heater and a temperature meter to keep track of it!


--------------------
“Everybody is a genius. But if you judge a fish by its ability to climb a tree, it will live its whole life believing that it is stupid.”
- Albert Einstein





Edited by Fluzzball (07/26/15 02:03 PM)


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InvisibleDr.Satan
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Registered: 06/26/15
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Re: Inoculation Question [Re: Fluzzball]
    #22000120 - 07/26/15 02:05 PM (8 years, 6 months ago)

Quote:

Fluzzball said:
Thanks everyone for your help! I will wait it out and see how things turn out. I inoculated 10 jars with the verm/brown rice flour method. Day 3 and still nothing. I am going to put them somewhere warmer, as they are only at about 70 degrees and I read keeping them at about 75-85 is an ideal temperature. Figured I'd find a bin, put some water in a bowl, use a fish tank heater and a temperature meter to keep track of it!




Oh day 3?? Don't worry about it man. Some of my BRF jars took over a week to begin to form mycelium. And now those same jars are doing so well that after they fully colonize I only consolidate them for a day or two before pins start to form causing me to have to begin to dunk and roll right away. 4 are fully done colonizing (two of which have already been dunked and rolled. And the rest are at over 95% colonization. I inoc'ed them on like June 28th or something like that and didn't see mycelium growth in any of them until like july 8th or around that time.

And remember patience is key. I was on like day 9 of no growth with these jars and I remember making a thread because I was worried I fucked up, but as it turns out I was just paying far to much attention to them while they were colonizing which made me worry over nothing lol. I believe the maximum time you should give them is 14 days, if you still see no white mycelium growing in the jars after 14 days then something went wrong.


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


Edited by Dr.Satan (07/26/15 02:20 PM)


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OfflinePsilosoulful

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Re: Inoculation Question [Re: Dr.Satan]
    #22000427 - 07/26/15 03:32 PM (8 years, 6 months ago)

Don't worry about the temp too much, brf jars will colonize fine in temps from 70-78f. Just leave it on a shelf and check on it in a couple of days. I've never used an incubator and don't think it's necessary. :twocents:


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OfflinetaGyo
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Re: Inoculation Question [Re: Psilosoulful]
    #22000444 - 07/26/15 03:38 PM (8 years, 6 months ago)

:whathesaid:


--------------------
Gyo's Better Grows
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OfflineFluzzball
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Re: Inoculation Question [Re: taGyo]
    #22001925 - 07/26/15 09:00 PM (8 years, 6 months ago)

Thank you everyone for your excellent feedback! It's greatly appreciated. I am just too darn excited so I feel like I am checking on them too much in anticipation! Haha! :tongue:


--------------------
“Everybody is a genius. But if you judge a fish by its ability to climb a tree, it will live its whole life believing that it is stupid.”
- Albert Einstein





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OfflinePsilosoulful

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Re: Inoculation Question [Re: Fluzzball]
    #22001973 - 07/26/15 09:07 PM (8 years, 6 months ago)

Quote:

Fluzzball said:
I am just too darn excited so I feel like I am checking on them too much in anticipation! Haha! :tongue:



We all know that feelin'! :lol:


Edited by Psilosoulful (07/26/15 09:08 PM)


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InvisibleDr.Satan
Mad Professor
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Registered: 06/26/15
Posts: 2,182
Re: Inoculation Question [Re: Psilosoulful]
    #22002779 - 07/27/15 03:12 AM (8 years, 6 months ago)

Quote:

Psilosoulful said:
Quote:

Fluzzball said:
I am just too darn excited so I feel like I am checking on them too much in anticipation! Haha! :tongue:



We all know that feelin'! :lol:



For sure! I'm looking at pins on one of my cakes like 8 times a day like *hmm I wonder if those grew any in the last 2 hours* haha. I can't help but constantly check on them as much as I can. Same with when I was growing weed, I always use to go outside and smoke bowls by my plants while I observed them. Good times. I love the processes of this sort of thing. I like watching things like bud and mushrooms grow from nothing it's so cool :laugh:


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


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OfflineFluzzball
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Re: Inoculation Question [Re: Dr.Satan]
    #22003575 - 07/27/15 09:37 AM (8 years, 6 months ago)

Haha yes! I feel oddly maternal over my shroom babies. O___O Well they say that plants grow better with positive vibes. Meh, who knows?! Hahaha! :crazy:


--------------------
“Everybody is a genius. But if you judge a fish by its ability to climb a tree, it will live its whole life believing that it is stupid.”
- Albert Einstein





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InvisibleMad Season
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Re: Inoculation Question [Re: Fluzzball]
    #22003662 - 07/27/15 09:56 AM (8 years, 6 months ago)

So it was warm to the touch? Not scalding hot? I've hot poured agar over spores, and they germinated just fine. Spores begin to die at 120 F. A temperature that's almost too hot to touch.


--------------------
contam and car window art
How to shroomery like a pro! (Seriously, everyone read this!)
Improve your sterile techniques! (A comprehensive guide to agar)
Links upon links of literally EVERYTHING UP TO DATE

AMU Q&A
No trees were killed in the sending of this message. However, a large number of electrons were terribly inconvenienced.


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OfflineFluzzball
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Re: Inoculation Question [Re: Mad Season]
    #22003707 - 07/27/15 10:09 AM (8 years, 6 months ago)

It was warm, but no it didn't make my fingertips hurt. I could hold it comfortably.

Here are photos. I inoculated Thursday night. There are 10 jars total.





--------------------
“Everybody is a genius. But if you judge a fish by its ability to climb a tree, it will live its whole life believing that it is stupid.”
- Albert Einstein





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OfflinetaGyo
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Re: Inoculation Question [Re: Mad Season]
    #22003719 - 07/27/15 10:11 AM (8 years, 6 months ago)

Quote:

Mad Season said:
So it was warm to the touch? Not scalding hot? I've hot poured agar over spores, and they germinated just fine. Spores begin to die at 120 F. A temperature that's almost too hot to touch.



:thumbup:


--------------------
Gyo's Better Grows
TNF Q&A :rockon:
AMU Q&A

Dominus fortunae meae sum


Edited by taGyo (07/27/15 10:12 AM)


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InvisibleDr.Satan
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Registered: 06/26/15
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Re: Inoculation Question [Re: Fluzzball]
    #22004340 - 07/27/15 12:44 PM (8 years, 6 months ago)

Quote:

Fluzzball said:
It was warm, but no it didn't make my fingertips hurt. I could hold it comfortably.

Here are photos. I inoculated Thursday night. There are 10 jars total.








Odds are they're fine then. Just give them their 14 days and if you reach the 14 day mark and still see no myc growing then you can chalk it up as a failure.


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InvisibleMad Season
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Re: Inoculation Question [Re: Dr.Satan]
    #22013827 - 07/29/15 08:50 AM (8 years, 6 months ago)

I must humbly disagree. I have a jar I inoculated with spores in April and it seems to have germinated mid July. If you've got enough jars, I'd just start up some more projects while waiting for those ones

Also seen stuff germinate on agar after a month or 2. :shrug: not sure why, but 2 weeks isn't anywhere near enough time to call it a failure. More like 1 or 2 months for me.


--------------------
contam and car window art
How to shroomery like a pro! (Seriously, everyone read this!)
Improve your sterile techniques! (A comprehensive guide to agar)
Links upon links of literally EVERYTHING UP TO DATE

AMU Q&A
No trees were killed in the sending of this message. However, a large number of electrons were terribly inconvenienced.


Edited by Mad Season (07/29/15 08:53 AM)


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OfflinePsilosoulful

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Re: Inoculation Question [Re: Mad Season]
    #22013859 - 07/29/15 08:57 AM (8 years, 6 months ago)

Quote:

Mad Season said:
I must humbly disagree. I have a jar I inoculated with spores in April and it seems to have germinated mid July.



:woah:


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InvisibleMad Season
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Re: Inoculation Question [Re: Psilosoulful]
    #22013881 - 07/29/15 09:05 AM (8 years, 6 months ago)

Ikr! I have just been thinking it was a failure, and had I them in the back of my closet, too lazy to clean them out. I checked the other day and found a pretty big round mass of tampanensis mycelium. If you've been following me around, you'll see I was bitching about my tamps not germinating on agar or grains after a month or 2. Well apparently I had to wait 3+ months LOL.

See: http://www.shroomery.org/forums/showflat.php/Number/21589847 those wbs jars are the ones that germinated lol

It goes to show, spores rarely are bunk. When you have such a massive population of them, you should still see something. I always can't help but wonder if people toss so many stuff with viable spores still in them, that they're just taking their time.


--------------------
contam and car window art
How to shroomery like a pro! (Seriously, everyone read this!)
Improve your sterile techniques! (A comprehensive guide to agar)
Links upon links of literally EVERYTHING UP TO DATE

AMU Q&A
No trees were killed in the sending of this message. However, a large number of electrons were terribly inconvenienced.


Edited by Mad Season (07/29/15 09:08 AM)


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OfflineNDStepp84
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Re: Inoculation Question [Re: Mad Season]
    #22013893 - 07/29/15 09:07 AM (8 years, 6 months ago)

IMO I would take that foil off, you don't need it with the dry verm layer. It may restrict air exchange or hold moisture in your dry verm layer increasing your chances of contamination.


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

"I am free, no matter what rules surround me. If I find them tolerable, I tolerate them; if I find them too obnoxious, I break them. I am free because I know that I alone am morally responsible for everything I do.
-Robert A. Heinlein 

:takingnotes: Links and teks:takingnotes:
ND's grow log and discussion
Plant thread


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OfflinetaGyo
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Re: Inoculation Question [Re: Mad Season]
    #22013954 - 07/29/15 09:23 AM (8 years, 6 months ago)

Quote:

NDStepp84 said:
IMO I would take that foil off, you don't need it with the dry verm layer. It may restrict air exchange or hold moisture in your dry verm layer increasing your chances of contamination.



:thumbup:
Quote:

Mad Season said:
Ikr!



:congrats:

I would have tossed that a long time ago.


--------------------
Gyo's Better Grows
TNF Q&A :rockon:
AMU Q&A

Dominus fortunae meae sum


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InvisibleMad Season
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Re: Inoculation Question [Re: taGyo]
    #22013988 - 07/29/15 09:30 AM (8 years, 6 months ago)

Haha yeah, I would have too. I've been too lazy, and forgot about them too. Thanks though :smile:. Pretty happy I didn't toss them :smile: I'm going to do some g2a soon, so I can finally have it on agar lol


--------------------
contam and car window art
How to shroomery like a pro! (Seriously, everyone read this!)
Improve your sterile techniques! (A comprehensive guide to agar)
Links upon links of literally EVERYTHING UP TO DATE

AMU Q&A
No trees were killed in the sending of this message. However, a large number of electrons were terribly inconvenienced.


Edited by Mad Season (07/29/15 09:34 AM)


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OfflineFluzzball
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Re: Inoculation Question [Re: NDStepp84]
    #22014128 - 07/29/15 10:02 AM (8 years, 6 months ago)

I ended up taking the foil off and putting mirco pore tape to see if that would speed up the process. So far out of 10 jars I've only had a single green mold contaminant(I took the lid off and noticed some accumulated moisture in the dry verm layer) and 4 of them have started to germinate and have healthy looking mycelium! :smile: I may be seeing smaller growth on the others but it's too soon to tell!

I've placed them on the hot water heater under a light towel and it's raised my temp from 70 to about 76.

I am way too excited!!


--------------------
“Everybody is a genius. But if you judge a fish by its ability to climb a tree, it will live its whole life believing that it is stupid.”
- Albert Einstein





Edited by Fluzzball (07/29/15 10:07 AM)


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OfflineNDStepp84
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Re: Inoculation Question [Re: Fluzzball]
    #22014252 - 07/29/15 10:30 AM (8 years, 6 months ago)

You don't need the tape over the holes, it will slow down colonization time.


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

"I am free, no matter what rules surround me. If I find them tolerable, I tolerate them; if I find them too obnoxious, I break them. I am free because I know that I alone am morally responsible for everything I do.
-Robert A. Heinlein 

:takingnotes: Links and teks:takingnotes:
ND's grow log and discussion
Plant thread


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OfflineFluzzball
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Re: Inoculation Question [Re: NDStepp84]
    #22015619 - 07/29/15 03:32 PM (8 years, 6 months ago)

Good to know! Thank you! :smile:


--------------------
“Everybody is a genius. But if you judge a fish by its ability to climb a tree, it will live its whole life believing that it is stupid.”
- Albert Einstein





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