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Offlinekalran
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I think I made a mistake
    #26356657 - 12/02/19 04:56 AM (4 years, 1 month ago)

Hello everyone!

I inoculated my grain-jars with Cube B+ about a 3 weeks ago.
The jars,so far, look like they are in a healthy condition, and I see new fresh mycelium every day.
In the process of learning how to do this (this is my first grow), I might have missed a step.

I did not take into account for gas-exchange when I prepared the jars, and they are now tightly sealed with no possibility for air- or gas-exchange.

- Does this ruin the whole batch?
- Is there a way to salvage this?

The jars have an air-lock seal underneath the lid. Can I remove the lid in a sterile glove-box, drill a hole in the lid, put on a filter-tape, and put it back on? And if so, approximately how large does the hole need to be in order for sufficient GE to be present?

Thanks in advance!


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Offlinehamonz
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Re: I think I made a mistake [Re: kalran]
    #26356668 - 12/02/19 05:14 AM (4 years, 1 month ago)

I would just leave it and start working on some more spawn to put to grains, if you fuck with it it'll just introduce contams.

Did you inoculate with a spore syringe or with agar?


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Offlinekalran
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Re: I think I made a mistake [Re: hamonz]
    #26356673 - 12/02/19 05:22 AM (4 years, 1 month ago)

I used a syringe. I'm all out of spores, and they are hard to get. I live in Norway which has strict laws against buying spores that lead to psychedelic mushrooms. I was nervous about buying them the first time + most spore-merchants require you to buy in bitcoin or certain alt-coins, which is a hassle for me.

Edit: And by the way I was asking if this was a legitimate way to salvage the operation. If I fuck it up, then yes, I fuck up. But as of now, the grains are already fucked if I don't allow GE, yes?


Edited by kalran (12/02/19 05:26 AM)


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Offlinecurious.psychonaut
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Re: I think I made a mistake [Re: kalran]
    #26356680 - 12/02/19 05:27 AM (4 years, 1 month ago)

I think they may stall without gas exchange (or worse, breed anaerobic bacteria). Before "fixing" anything, you might want to wait for the myc to actually stop growing.

Two ways to add GE come to mind:
1. Unscrew lids a bit & hope for the best.
2. Prepare new lids with proper filter, PC them in foil & bring the closed foil into your SAB (still air box = glove box without gloves). Unscrew old lids, screw on new ones.

One hole punched with a standard philips/pozidriv #2 screwdriver (5-6mm) is all that's needed. Cover with micropore tape or stuff with poly-fil.

Edit: just noticed you are talking grain jars, so removed the stuff about BRF.


--------------------
My LAGM2020 grow log



Edited by curious.psychonaut (12/02/19 05:36 AM)


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Offlinekalran
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Re: I think I made a mistake [Re: curious.psychonaut]
    #26356683 - 12/02/19 05:32 AM (4 years, 1 month ago)

Quote:

curious.psychonaut said:
Do your jars have a dry verm layer on top?




No. I used a TEK that did not explicitly describe the use of vermiculite, so I ditched it.

I feel like pre-cooking the lids is a safe way of doing this. I will try this and hope for the best! Thanks a lot!

Edit: Any particular reason I should wait for the mycelium to stall before I attempt this? Just curious :smile:


Edited by kalran (12/02/19 05:34 AM)


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Offlinecurious.psychonaut
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Re: I think I made a mistake [Re: kalran]
    #26356691 - 12/02/19 05:41 AM (4 years, 1 month ago)

Quote:

kalran said:
No. I used a TEK that did not explicitly describe the use of vermiculite, so I ditched it.



Sorry, I got mixed up, it's grain jars, so you don't have dry verm on top--no TEK calls for that (AFAIK).

Quote:


Edit: Any particular reason I should wait for the mycelium to stall before I attempt this? Just curious :smile:



Because there's always a chance to contaminate, so why risk it if it still works?


--------------------
My LAGM2020 grow log



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OfflineMycoactive
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Re: I think I made a mistake [Re: kalran]
    #26356693 - 12/02/19 05:41 AM (4 years, 1 month ago)

Quote:

kalran said:
Quote:

curious.psychonaut said:
Do your jars have a dry verm layer on top?




No. I used a TEK that did not explicitly describe the use of vermiculite, so I ditched it.

I feel like pre-cooking the lids is a safe way of doing this. I will try this and hope for the best! Thanks a lot!

Edit: Any particular reason I should wait for the mycelium to stall before I attempt this? Just curious :smile:




The farther along the mycelium is, the less of a chance competing microorganisms have a chance to take hold. In a sterile environment and after sterilizing the outside of your jars, you can quickly loosen the lid and then retighten. Gas exchange happens fairly rapidly if there's a large buildup of CO2 inside the jars so there's no need to keep the lids loose. You might have to do this again if the myc stalls again, but it's safer than maintaining loose lids.


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InvisibleSpunkyMonkey88
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Re: I think I made a mistake [Re: curious.psychonaut]
    #26356694 - 12/02/19 05:42 AM (4 years, 1 month ago)

Quote:

curious.psychonaut said:
I think they may stall without gas exchange (or worse, breed anaerobic bacteria). Before "fixing" anything, you might want to wait for the myc to actually stop growing.

Two ways to add GE come to mind:
1. Unscrew lids a bit & hope for the best.
2. Prepare new lids with proper filter, PC them in foil & bring the closed foil into your SAB (still air box = glove box without gloves). Unscrew old lids, screw on new ones.

One hole punched with a standard philips/pozidriv #2 screwdriver (5-6mm) is all that's needed. Cover with micropore tape or stuff with poly-fil.

Edit: just noticed you are talking grain jars, so removed the stuff about BRF.




A canning lid will still seal even without the ring in at all... I'd just let them go until they stop, shake them up and do a g2g to some jars with a filter on them, right away...

Itll be slow to colonize but granted that syringe wasn't super bacterial it would work...

I'm assuming he doesn't want to work with agar which would be the best chance to save his jars.

If he wants to do agar I would let the jars grow until he can pluck a kernel with mycelium on it and put it on a plate and transfer until clea .


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OfflineA.k.aM
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Re: I think I made a mistake [Re: curious.psychonaut]
    #26356695 - 12/02/19 05:43 AM (4 years, 1 month ago)

Quote:

curious.psychonaut said:
I think they may stall without gas exchange (or worse, breed anaerobic bacteria). Before "fixing" anything, you might want to wait for the myc to actually stop growing.

Two ways to add GE come to mind:
1. Unscrew lids a bit & hope for the best.
2. Prepare new lids with proper filter, PC them in foil & bring the closed foil into your SAB (still air box = glove box without gloves). Unscrew old lids, screw on new ones.

One hole punched with a standard philips/pozidriv #2 screwdriver (5-6mm) is all that's needed. Cover with micropore tape or stuff with poly-fil.

Edit: just noticed you are talking grain jars, so removed the stuff about BRF.





Exactly what I was going to say. Wait to see if it stalls and if it does SAB new sterile lids. Idk if you have tyvek at your post offices, but the ONLY jar I’ve had contaminate beyond use was the only one where I used Micropore tape instead of tyvek with silicon SHIP on top.


--------------------
LAGM2020


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InvisibleSpunkyMonkey88
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Re: I think I made a mistake [Re: SpunkyMonkey88]
    #26356697 - 12/02/19 05:45 AM (4 years, 1 month ago)

Or you could put some lids with filters in a pp5 container or a grow bag and PC them then put then replace the other lids with those in a SAB..


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Offlinekalran
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Posts: 31
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Re: I think I made a mistake [Re: A.k.a]
    #26356718 - 12/02/19 06:06 AM (4 years, 1 month ago)

Quote:

Exactly what I was going to say. Wait to see if it stalls and if it does SAB new sterile lids. Idk if you have tyvek at your post offices, but the ONLY jar I’ve had contaminate beyond use was the only one where I used Micropore tape instead of tyvek with silicon SHIP on top.




Forgive my noob-ness, but what is "SAB", and isn't tyvek-tape air- and water-tight, defeating the purpose? :confused:
I might get my hands on some tyvek, I think. Does the tape need to be sterilized as well, and if so, how do I go about that?
And "silicon SHIP"? New terms to me :frown:

EDIT: Ah, SAB = Still Air-Box - sorry I'm dumb :blush:


Edited by kalran (12/02/19 06:15 AM)


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Offlinekalran
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Re: I think I made a mistake [Re: SpunkyMonkey88]
    #26356725 - 12/02/19 06:12 AM (4 years, 1 month ago)

Quote:

SpunkyMonkey88 said:
I'm assuming he doesn't want to work with agar which would be the best chance to save his jars.

If he wants to do agar I would let the jars grow until he can pluck a kernel with mycelium on it and put it on a plate and transfer until clea .




I only discovered the features of agar after I inoculated the jars with the syringe, and all the tutorials I looked at on YT used syringes.

And when "cleaning" the transfer with agar-dish to agar-dish(?), approximately how many transfer is necessary to achieve completely clean mycelium?


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InvisibleSpunkyMonkey88
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Re: I think I made a mistake [Re: kalran]
    #26356727 - 12/02/19 06:13 AM (4 years, 1 month ago)

Quote:

kalran said:
Quote:

Exactly what I was going to say. Wait to see if it stalls and if it does SAB new sterile lids. Idk if you have tyvek at your post offices, but the ONLY jar I’ve had contaminate beyond use was the only one where I used Micropore tape instead of tyvek with silicon SHIP on top.




Forgive my noob-ness, but what is "SAB", and isn't tyvek-tape air- and water-tight, defeating the purpose? :confused:
I might get my hands on some tyvek, I think. Does the tape need to be sterilized as well, and if so, how do I go about that?
And "silicon SHIP"? New terms to me :frown:




A SAB is a still air box, and it is crucial for success with this hobby. It's just a rubbermaid tub with holes for your arms.  You can open sterile jars and such inside with out contaminating them...look it up..


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InvisibleSpunkyMonkey88
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Re: I think I made a mistake [Re: kalran]
    #26356731 - 12/02/19 06:16 AM (4 years, 1 month ago)

Quote:

kalran said:
Quote:

SpunkyMonkey88 said:
I'm assuming he doesn't want to work with agar which would be the best chance to save his jars.

If he wants to do agar I would let the jars grow until he can pluck a kernel with mycelium on it and put it on a plate and transfer until clea .




I only discovered the features of agar after I inoculated the jars with the syringe, and all the tutorials I looked at on YT used syringes.

And when "cleaning" the transfer with agar-dish to agar-dish(?), approximately how many transfer is necessary to achieve completely clean mycelium?




Ot depends but on how dirty your initial transfer is but I did this with a pretty bacterial jar and 3 transfers later its clean as far from what the (my) eye can tell.

This would be your best chance at saving your culture. If you do this all right you wont need to ever buy spores again..


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Offlinekalran
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Re: I think I made a mistake [Re: SpunkyMonkey88]
    #26356734 - 12/02/19 06:20 AM (4 years, 1 month ago)

Quote:

SpunkyMonkey88 said:
This would be your best chance at saving your culture. If you do this all right you wont need to ever buy spores again..




I will definitely prepare some agar-dishes now. If they can actually save a bacterial-infected culture??? Man, then it's magic! How long can you store a cultivated agar-dish?

Appreciate the help man! :thumbup:

EDIT: And btw, what kind of tyvek-tape should I be looking for?
Are there any other good alternatives? I've read more up on micropore tape from 3M, and many reports indicate that the tape is bad.

EDIT 2: So I figured out what Tyvek was, and what I'm going to use.
I can use paper cut out from tyvek-envelopes, which they actually have at my local store :crazy2:


Edited by kalran (12/02/19 07:27 AM)


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Offlinecurious.psychonaut
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Re: I think I made a mistake [Re: kalran]
    #26356800 - 12/02/19 07:41 AM (4 years, 1 month ago)

Quote:

kalran said:
I will definitely prepare some agar-dishes now. If they can actually save a bacterial-infected culture??? Man, then it's magic! How long can you store a cultivated agar-dish?




Best decision :awesome:! You can store a dish for months in the fridge (myc stops growing). You can also make a slant (test tube with slanted agar & wood) and it can last for years.

Quote:

EDIT: And btw, what kind of tyvek-tape should I be looking for?
Are there any other good alternatives? I've read more up on micropore tape from 3M, and many reports indicate that the tape is bad.



The most common lids are, in order of filtering quality & as far as I understand:
1. synthetic filter disc (~.2 microns)
1. cellulose filter disc (thicker and has to be kept dry so maybe not as practical)
3. poly-fil (has to be really tight: I fold and twist the shit out of it & use tweezers from the other side to get as much as I can and then pull with fingers)
4. micropore (it's just a bandage, not really a filter, but works acc/to many, my experience is also good so far (only 8 jars though))
4. tyvek (advertized as 1 micron, some bacteria are smaller)
6. normal lid, unscrewed a bit (no filtration but can work: a bit like a petri which also works, even if unwrapped).

Check the search results for TEKs on these variations.


--------------------
My LAGM2020 grow log



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