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OfflineMigraine
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Spores to LC
    #11239292 - 10/13/09 02:40 PM (8 years, 1 month ago)

I received a spore print and I don't know how clean it is.

I have made 5 agar dishes from the spores with the intention for going to grain if no contams.

I have just made an LC solution which I would like to also use with the spores.

My problem is - how best to move spores into the LC without bringing any contams.

#1.  Hold print over LC jar and scrape.

#2.  Scrape spores into a small shot glass containing low percentage hydrogen peroxide - then transfer that solution into the LC.

#3.  Sterile water puddle on print and move solution to LC.

Any thoughts would be helpful - contams love LC ... I want to keep them out.

Thanks


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Invisiblemyco_mikey_mo
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Re: Spores to LC [Re: Migraine]
    #11239346 - 10/13/09 02:49 PM (8 years, 1 month ago)

H2O2 kills spores.


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Invisiblemyco_mikey_mo
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Re: Spores to LC [Re: myco_mikey_mo]
    #11239435 - 10/13/09 03:07 PM (8 years, 1 month ago)

and i would just reccomend streaking the spores with an inoculation loop or something like that. honestly though i have always just made a syringe and squirted some solution in my LC. works great for me. also a glovebox or flow hood would really help


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Offlinesolumvita
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Re: Spores to LC [Re: myco_mikey_mo]
    #11239467 - 10/13/09 03:14 PM (8 years, 1 month ago)

wait until the spores on the plates have germinated and are visibly clear from contams and then transfer a piece from a clean plate into the LC broth.


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OfflineMigraine
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Re: Spores to LC [Re: solumvita]
    #11240080 - 10/13/09 04:59 PM (8 years, 1 month ago)

Thanks for the replies.

I do have and use a flow hood.

I wanted to run a fast path so I scraped and dropped spores directly into the LC.

Contam - time will tell.

I will do one more from my agar when they grow out.

Thanks all.


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Invisible13shroomsM
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Re: Spores to LC [Re: Migraine]
    #11246400 - 10/14/09 04:26 PM (8 years, 1 month ago)



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OfflinePuma
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Re: Spores to LC [Re: 13shrooms]
    #11444627 - 11/12/09 11:33 PM (8 years, 11 days ago)

Well, how did it turn out? You've kept us in suspense long enough...!

Out with it!

Photos? What's the status now of that grow?


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Invisible13shroomsM
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Re: Spores to LC [Re: Puma]
    #11444686 - 11/12/09 11:40 PM (8 years, 11 days ago)

:lmafo:


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OfflineMigraine
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Re: Spores to LC [Re: 13shrooms]
    #11453473 - 11/14/09 02:13 PM (8 years, 9 days ago)

Sorry for delay - The spore print was from a Golden Teacher.

The five quart jars of rye - from agar dishes are only at 70% colonized - so I have not taken them to substrate yet.

I dropped spores into a 4% light malt LC solution.  After 20 days small islands of mycelium started forming on the surface.  I placed the LC on a magnetic stirrer for two minutes every day.  I have placed the LC in the frig - may not use it. In Aug I made a similar LC of King Oyster and Lions Mane.  I had to leave home for a month - so no stiiring.  Before I left I had the same formation taking place - I though it was contam - that is why I just left it.  To my suprise, when I returned, I found a three inch thick mycelium slab sitting on the surface of the LC.


My goal now will be to fruit from the rye jars and clone to agar from a nice looking mushroom.  Once I know I have a contam free dish - I will take it to slants (for my library) and then a few small rye jars.  Once those rye jars have colonized, I will extract mycelium from one and use IT as my LC master as Agar outlined in one of his posts.

I have read almost every one of Agar's threads - I don't know how I missed that one - Thanks 13shrooms for pointing it out.

Pics when I have something interesting.


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InvisibleBlimeyGrimey
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Re: Spores to LC [Re: 13shrooms]
    #11459904 - 11/15/09 03:13 PM (8 years, 8 days ago)



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OfflinePuma
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Re: Spores to LC [Re: BlimeyGrimey]
    #11464355 - 11/16/09 01:27 AM (8 years, 8 days ago)

Interesting update. Thanks for that I guess maybe spore-to-LC isn't really the fast path after all. I thought it would be too, but after ten days I'm seeing very little formation, other than a few small squiggles, and a sort of white film along the inside edge of the jar, at the surface of the liquid. I wonder if it's definitely growth, possibly contam, or just residue of malt extract. Hopefully it'll soon be more obvious.

That's quite something, the puck of mycelium from those jars you left!

It sounds like agar media and grain might be preferable to dropping spores in liquid solution. Although I spent all my Orissa spores on the LC attempt (so I'm hoping it pans out) I'll make LC starting with another strain, using RR's preferred technique of adding sterile water to a colonized grain jar, shaking it up, and charging syringes with the resulting myc-rich water. It seems like that's the best way to speed up the time (as grains colonize faster than water) as well as minimizing the odds of contam, as you can inspect visually... as others here have noted.

Best of luck with your cultures. I would be tempted to drop some of that LC in your fridge into some agar to see if its clean.


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InvisibleBlimeyGrimey
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Re: Spores to LC [Re: Puma]
    #11465194 - 11/16/09 06:30 AM (8 years, 8 days ago)

Quote:

Puma said:
I thought it would be too, but after ten days I'm seeing very little formation, other than a few small squiggles, and a sort of white film along the inside edge of the jar, at the surface of the liquid.




Sounds like bacterial contamination.


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OfflinePuma
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Re: Spores to LC [Re: BlimeyGrimey]
    #11465235 - 11/16/09 06:56 AM (8 years, 8 days ago)

Uh oh.....

Here's what it looks like. I'm clinging to the hope that it might be mycelium growing up there because that's where there's access to oxygen (although I've been shaking too). The white gummy stuff flakes off sometimes when shaken.







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