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OfflineOPB
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Registered: 05/30/15
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Sustainable Sclerotia - Long Term Cultures
    #21877463 - 06/30/15 10:36 AM (8 years, 6 months ago)

I was reading this very informative post about long term storage of mycelium in distilled water.

http://www.shroomery.org/8509/Resurrecting-a-Better-Method-for-Long-Term-Storage-of-Mushroom-Cultures

I would like to be able to store some sort of truffle producing material long term so that I wouldn't need to order more spores.

Could I simply take a colonized jar (sfd with self healing port) and inject sterlized water (as one would when making a syringe for water suspension transfers of mycelium) then draw the water back out, dilute, and then store? It's seems that I should be able to but I worry that the distilled water will pick up nutrients from the grains and prevent the stasis which is achieved with the previously mentioned distilled water suspension technique.

Assuming this works I would transfer this to agar before using as a inoculant to avoid contams.

Think this should work? Are there better options avaliable to a quick learning noob?


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InvisibleFriedEggS
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Registered: 09/22/14
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Re: Sustainable Sclerotia - Long Term Cultures [Re: OPB]
    #21877466 - 06/30/15 10:37 AM (8 years, 6 months ago)

I think you'd be better off using slants for long term storage (stored in the refrigerator)


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OfflinePsilosopherr
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Re: Sustainable Sclerotia - Long Term Cultures [Re: FriedEgg]
    #21877544 - 06/30/15 10:59 AM (8 years, 6 months ago)

yeah slants are the go-to 'long term storage tek.'

I think you can find a write-up of how to make slants in my signature: under 'how frank gets shit done'

edit: fuck it I'll link you anyway
http://www.shroomery.org/forums/showflat.php/Number/18182209#18182209


Edited by Psilosopherr (06/30/15 10:59 AM)


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InvisibleMad Season
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Re: Sustainable Sclerotia - Long Term Cultures [Re: Psilosopherr]
    #21877560 - 06/30/15 11:05 AM (8 years, 6 months ago)

If you are able to do agar work but don't have any tubes or small containers for easy storage of agar, there's always LC/LI and syringes. I know a certain TC used a 2007 oyster LC, and it worked like a charm.

You just gotta make sure to test your syringes on a few plates of agar to ensure they're clean.


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OfflineOPB
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Re: Sustainable Sclerotia - Long Term Cultures [Re: Mad Season]
    #21877583 - 06/30/15 11:10 AM (8 years, 6 months ago)

I will look into slants. Thanks.

Quote:

Mad Season said:
If you are able to do agar work but don't have any tubes or small containers for easy storage of agar, there's always LC/LI and syringes. I know a certain TC used a 2007 oyster LC, and it worked like a charm.

You just gotta make sure to test your syringes on a few plates of agar to ensure they're clean.




This basically is a liquid culture in my understanding. The difference is just that there are no nutrients and eventually no oxygen so the culture is basically zombified. If my understanding is correct one advantage this would have over traditional liquid culture is that the lack of nutrients also prevents contams.


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OfflinePsilosopherr
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Re: Sustainable Sclerotia - Long Term Cultures [Re: OPB]
    #21877597 - 06/30/15 11:13 AM (8 years, 6 months ago)

are their two versions of LC or something? I always thought TC's avoided it but now I see more and more of them talking about it


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OfflineOPB
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Re: Sustainable Sclerotia - Long Term Cultures [Re: Psilosopherr]
    #21877629 - 06/30/15 11:24 AM (8 years, 6 months ago)

Quote:

rbalzer said:
are their two versions of LC or something? I always thought TC's avoided it but now I see more and more of them talking about it




Some swear by it, some swear about it. The key though, in my understanding, is that the problems arise when multiplying cultures in the nutrient rich solution. This technique uses distilled water which is about as lacking in nutrients as physically possible. The point is to capture the material and hold it in suspended animation. My main concern is the potential for the grain nutrients to stop this from working. If the technique I previously described works then it would be by far my easiest and most sterile option. According to the resource I linked to in the original post you could be looking at 5-10 years of viability at room temp!


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OfflinePsilosopherr
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Re: Sustainable Sclerotia - Long Term Cultures [Re: OPB]
    #21877639 - 06/30/15 11:26 AM (8 years, 6 months ago)

Quote:

OPB said:
Quote:

rbalzer said:
are their two versions of LC or something? I always thought TC's avoided it but now I see more and more of them talking about it




Some swear by it, some swear about it. The key though, in my understanding, is that the problems arise when multiplying cultures in the nutrient rich solution. This technique uses distilled water which is about as lacking in nutrients as physically possible. The point is to capture the material and hold it in suspended animation. My main concern is the potential for the grain nutrients to stop this from working. If the technique I previously described works then it would be by far my easiest and most sterile option. According to the resource I linked to in the original post you could be looking at 5-10 years of viability at room temp!



interesting :thumbup:

some swear by it, others swear about it, very clever :lol:


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InvisibleMad Season
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Re: Sustainable Sclerotia - Long Term Cultures [Re: Psilosopherr]
    #21877641 - 06/30/15 11:26 AM (8 years, 6 months ago)

Kind of. LC is obvious. Nutrients and water for mycelium to colonize. Then put it in a syringe. The problem with LC is it has so many nutrients, it can contaminate with a VERY low contaminate spore count.

LI is you have a colonized dish of agar, add sterile water and blend it up together. Way less nutrients are in this solution.

Although both can contaminate, I find LI to have a less chance. However I still seriously recommend testing both on agar dishes to ensure they're clean. Agar tested liquids are generally clean af.


--------------------
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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OfflinePsilosopherr
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Re: Sustainable Sclerotia - Long Term Cultures [Re: Mad Season]
    #21877767 - 06/30/15 11:59 AM (8 years, 6 months ago)

Quote:

Mad Season said:
Kind of. LC is obvious. Nutrients and water for mycelium to colonize. Then put it in a syringe. The problem with LC is it has so many nutrients, it can contaminate with a VERY low contaminate spore count.

LI is you have a colonized dish of agar, add sterile water and blend it up together. Way less nutrients are in this solution.

Although both can contaminate, I find LI to have a less chance. However I still seriously recommend testing both on agar dishes to ensure they're clean. Agar tested liquids are generally clean af.



ah that makes sense.

Thank you both for clarifying. (sorry for thread jacking :smile:)


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OfflineOPB
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Re: Sustainable Sclerotia - Long Term Cultures [Re: Mad Season]
    #21877822 - 06/30/15 12:09 PM (8 years, 6 months ago)

I guess I can always just try it and if it doesn't work have a backup plan. If anyone has any other input I'm all ears. Otherwise I'll give it a go and see what's happens!


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Invisiblegrainbrain
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Re: Sustainable Sclerotia - Long Term Cultures [Re: OPB]
    #21878072 - 06/30/15 01:11 PM (8 years, 6 months ago)

I prefer an ALC agar liquid culture to a GLC grain liquid culture.

Drop a clean wedge into sterilized water, let grow out for two weeks then put into cold storage.  This gives you both ready to use inoculant and long term storage.  I have an ATL7 culture that is 3+ years old, still works great.


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OfflineOPB
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Re: Sustainable Sclerotia - Long Term Cultures [Re: grainbrain]
    #21878132 - 06/30/15 01:27 PM (8 years, 6 months ago)

Quote:

grainbrain said:
I prefer an ALC agar liquid culture to a GLC grain liquid culture.

Drop a clean wedge into sterilized water, let grow out for two weeks then put into cold storage.  This gives you both ready to use inoculant and long term storage.  I have an ATL7 culture that is 3+ years old, still works great.




I will have to give that a try as well. I haven't experimented with agar just yet but it is on my short list of tools to learn. I just like this idea because of its simplicity and sterility. If you start with a clean colonized jar and sterlized water it seems nearly foolproof. You don't even need to use your SAB, just flame sterilize each step and keep the ports clean. It seems like the best plan would be to store using a couple of methods for a bit of healthy redundancy.


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