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Mushrooms, Mycology and Psychedelics > Mushroom Cultivation
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Re: Liquid culture [Re: Quwo]
    #27599241 -

Hahaha

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Re: Liquid culture [Re: Notarealguy] * 2
    #27599312 -

I am late to join the party. The lc jar and those bags are contaminated. Cheers for trying anyway. There has only been 1 company that could consistently make spore syringes that were clean enough for lc's. That was PF. His syringes were always super viable and clean. Don't know how he did it some consistently.
As for making lc's with spores, its gonna be very rare to do so successfully.
I use to grow out some spores on a petri, make a transfer to another petri once they were an inch wide of what appeared to be healthy mycelium, then once that second plate got about an inch or more of growth, I would get a syringe full of sterile water with a fat 14-16g needle on it, in a sterile glove box or hood I would open the pertri, cut up a pea to marble size area of the mycelium at the edge of the new growth using the tip of the needle to shred the mycelium, criss cross scraping, doing little circles, then squirt a few drops of water on it from the same syringe, then suck it up. Then I would use that syringe to inoc a lc along with a few 1/2 gallon jars of grain. Let them both grow for a week or 2. If the grain spawn looked good and if the lc looked good I would use the lc for some more jars. Easy peasy.
A health lc will look wispy. The mycelium will grow and grab onto the other mycelium. Yours isolated into little balls. A bad sign. Although your lc stayed clear. A milky lc is a sure sign of a yeast or something stinky.


--------------------
Stop experimenting half way through your first grow. Grow it to maturity, watch it, learn from it. Do this a few times then experiment with different ideas and figure out what works best for you.


My Legacy
https://www.shroomery.org/forums/showflat.php/Number/22140987#22140987

Teh=The
I need to proofread

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Re: Liquid culture [Re: tahoe]
    #27599320 -

If I remember right from someone who claimed to know him he took a short print to get the junk out and then printed in a Sterile jar with the cap suspended on a metal mesh. Either way spores to lc is like playing roulette you'll win big or have a high probability to lose it.

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Re: Liquid culture [Re: tahoe]
    #27599419 -

I noticed some growth starting on the Petri dishes yesterday. Sorry for the quality, I’ll go back today and take better pics.

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Re: Liquid culture [Re: Quwo]
    #27600152 -

Looks good so far, but next time you should use just a drop per petri dish.


It'll be another day or two before you'll be able to tell if it's any good. I'm guessing there will be some contamination that appears on your dishes along with the mycelium that's developing.


You would probably be best off taking a transfer from the healthiest part, let it grow out on another dish, and make a new LC from that.

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Re: Liquid culture [Re: Notarealguy]
    #27600249 -

Yea, I plan on doing the needle poke method. I’m wondering if I can use that method and inject through a self healing port.

And yeah, I was going for a drop per dish, but in all honesty, that was actually my first time ever working with Petri dishes or agar, so it was a little challenging getting the feel for it and dropping a single drop my first time. I should’ve practiced with water first. But we live and we learn, right?

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Re: Liquid culture [Re: Quwo]
    #27600314 -

I did the same thing the first time I put LC onto agar.  Forgot to break the plunger seal and sprayed a tenth of the syringe in there haha.


The poke method works through a SHIP:thumbup: 

just make sure to leave the foil on your LC jar until you inject in the SAB.  Otherwise you'll have to wipe it with an alcohol rag, and that can contam your LC because your needle is potentially dragging in contams that were clinging to the rag or paper towel.


Also make sure to do at least one more transfer to a new plate before you make the LC. I wouldn't use your test plate for the needle poke. 

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Re: Liquid culture [Re: Notarealguy]
    #27600519 -

I would recommend doing the poke from the dirty LC plate, or preferably a new germ plate. Taking that to a BRF puck, then poking a sample from the edge of that culture to transfer to a new agar plate. Let that transfer grow out a bit, then take a poke from the cleanest looking leading edge, and use that to inoculate a batch of LC. Test that LC in a small jar of grains or a plate, and if it’s good, then use that LC and the other clean agar wedges to inoculate your grain bags. Easy peasy lemon squeezy.


--------------------
Two eyes to look, One eye to see.
Prying open my third eye

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Re: Liquid culture [Re: Notarealguy]
    #27600852 -

Can I can do the first transfer to another plate using the poke method too?

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Re: Liquid culture [Re: Quwo]
    #27601174 -

Yeah no problem, it'll do the same thing a scalpel would do.    When you do the poke just poke it one time You don't need to squirt any water out or scrape at it, just one poke, then pop out the agar plug from the needle onto your new agar plate.


Even if you can't see any mycelium, it's likely still there.  It'll take a couple days to see any growth with such a small piece of tissue. But that's no big deal.



Honestly I would recommend just using a scalpel or any small knife you can get red hot for your first transfer. It'll be easier to deal with than the syringe IMO.

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Re: Liquid culture [Re: Notarealguy]
    #27612730 -

UPDATE
After transferring some of the questionable LC on some
Agar. How does it look guys? When should I transfer? Is there enough mycelium to do the needle poke method?


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Re: Liquid culture [Re: Quwo] * 1
    #27612741 -

Looks dirty as fuck. Not questionable LC anymore, lol, there is no question. You can try the poke. Do so sooner rather than later. I imagine that technique is not as effective against sporulating mold instead of bacteria, but it is worth a try. Be prepared to take lots of follow up transfers to clean it up. Or just start over from spore.m in agar.


--------------------
Two eyes to look, One eye to see.
Prying open my third eye

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Re: Liquid culture [Re: MysticMycologist] * 1
    #27612779 -

Probably will. I still want to try to clean this up because I’m out of spores for this strain.

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Re: Liquid culture [Re: Quwo] * 2
    #27612809 -

All successful growers had to go through a time of learning good sterile work habits.

For you to bring those pictures to your thread, tells me you should reexamine your sterile techniques.

My suggestion to you would be Is, forget lc's for awhile and work on being successful with spores to agar then agar to grain and grain spawn to your substrate..


--------------------
     

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Re: Liquid culture [Re: Quwo] * 1
    #27612832 -

Quwo said:
Probably will. I still want to try to clean this up because I’m out of spores for this strain.


I wouldn't try cleaning up a LC contaminated with mold. The mold and mushroom mycel will be well entwined. Buying new spores of this strain
or waiting until you get access to the marketplace would be the better move.


--------------------
🍄PANAEOLUS IN MONOTUBS🍄
🍄OFFICIAL SEMILANCEATA THREAD🍄



⭐Baba's Write Ups and Link Collection⭐

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Re: Liquid culture [Re: Baba Yaga]
    #27612995 -

Yup start fresh from spores.  There literally no salvation for a single one of those dishes.

LC is something you should try once you have a good grasp on agar.


Just use some spores an agar, get a grow outta that, select a few clones from the MS grow and run those next.  It's super straightforward.

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Mushrooms, Mycology and Psychedelics > Mushroom Cultivation
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