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Liquid Culture
    #27582768 -

This is my first time using liquid culture to expand a spore syringe for bulk use. I understand you can only see so much visually when it comes to LC. I’m asking for advice though because this is my first time and I really don’t know what to be looking for. Below I’ve attached some photos from day 4 after inoculating. The jars have been incubating at 82 degrees , and I’ve been mixing them once daily since the first mixing 48 hours after inoculation. The pics are before and after mixing. Please let me know what you think.

Edited by Quwo (12/17/21 09:32 PM)

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Re: Liquid culture [Re: Quwo]
    #27582772 -

That doesn't look too good but I'm not an expert so I'd wait for someone who is. However, it is generally discouraged to put spore syringes to LC due to their inherent dirtyness. It's better to use agar.

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Re: Liquid culture [Re: Quwo]
    #27582823 -

If your making LC from spores, you need to use agar to clean up your spores first.

  Shooting spore solution into LC fails most of the time. Because spore solution isn't fully sterile, it contains some contamination practically every time.  Once the contam hits the sterile LC it's game over.


So if you don't want to waste your time and get frustrated, get a small still air box, and some agar equipment.


82f is a little too warm, try to shoot for 70-72f.

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Re: Liquid culture [Re: Notarealguy] * 2
    #27582830 -

I would never use spore LC, especially if it's been cooking at 82. Agar, then LC, never the other way around.

Incoming: people for whom it worked once.


--------------------
* Smarts videos :teacher:
* :thumbup: Planet of the APES:thumbup:
 
I'm a fungal white supremacist.

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Re: Liquid culture [Re: Smartattack]
    #27583011 -

This is a good lesson to learn early. A spore syringe by definition of the way it's made is not sterile. I tried taking a shortcut this way thinking that vendor syringes should be clean enough but I just ended up with very bacterial spawn. Need to go to agar first and transfer to get clean isolate for LC

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Re: Liquid culture [Re: Notarealguy]
    #27583052 -

Yea I understand it’s not sterile and has higher risk for contamination. I was only doing it so I don’t need to buy a bunch of spore syringes, as I’m going to be inoculating 100 pounds of grain spawn bags. As for the temperature… I thought it was too high as well, I was just following the instructions that came with the LC jars I got from midwestgrowkits. The instructions say to keep the LC from 80-85 while it’s incubating… I thought 70-72 is for fruiting. All that aside I’m fully aware of the risks. I just want to know how it’s looking so far. I was probably going to test shooting up one bag first after mycelium has grown. Thanks for the help!

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Re: Liquid culture [Re: Quwo] * 1
    #27583076 -

The problem is, no matter how experienced a grower, nobody can say a single thing about how good it is. They are lying if they say they can. This is whole point of agar, so you can actually test it and see. Be aware that you are very likely sending 100 lbs of grain straight to compost. By the sound of it you are also buying pre made spawn bags, this will be a fairly pricey fail in money and time.


--------------------
* Smarts videos :teacher:
* :thumbup: Planet of the APES:thumbup:
 
I'm a fungal white supremacist.

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Re: Liquid culture [Re: Smartattack]
    #27583081 -

All else aside, do these jars look contaminated or not?

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Re: Liquid culture [Re: Quwo]
    #27583092 -

It's not really possible to tell man, they aren't cloudy, so that's good I suppose.


If your really going to do a 100 pounds of grain spawn, you're really taking a big risk by not using agar.


The time and effort it might save you will bite you in the ass.  I've done large grows with spores, when I first started.  It works until it doesn't.  And when it fails and all your money and work is wasted, it's a horrible feeling.

Agar seems complicated, but it's really not.  Check out pasty plates


You only need one drop of spores, you don't need to buy anymore than you have.

When you make a LC from spores you should use the absolute minimum amount of spore solution as possible to limit the potential of a contam getting in.  I'm guessing you used a decent amount to get that much growth so quickly.    I think you'll get way more weight in the end if you go the agar route:thumbup:

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Re: Liquid culture [Re: Quwo]
    #27583192 -

Quwo said:
All else aside, do these jars look contaminated or not?


No way to tell, man. When a culture grows out on agar, it gets a fixed shape which helps.  In LC, that doesn't happen.


--------------------
Sadly remembering when the Shroomery wasn't a closed minded mutual admiration society for OCD cargo cultists.  RIP methodical science.

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Re: Liquid culture [Re: OldManRiver]
    #27583203 -

I'm not an expert but I do make a good amount of lc. I can't say with 100% certainty but I'd chuck it in the toilet.

I don't like the way the myc looks.

Also good luck I would never give spore to lc of you are gonna make an investment in spawn bags you might wanna invest in some agar plates and a sab

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Re: Liquid culture [Re: Drboomer]
    #27583287 -

It looks uniform and not cloudy at least, you might be alright.

The growth seems a bit shredded for no stir bar but maybe you are able to shake the fuck out of it against that sharp metal on the lid.

Edit; oh now I see the bar lol.

Double edit: I mean 4 days is pretty fast to get that much growth in the LC from spore I dunno man. I wouldn't put it on 100lbs of grains.


--------------------
- Sandbag Tek - How To Sterilize Spawn Bags - All About Static Pressure / Pressure Drop for DIY Flow Hoods - Sandman's LC Tek-

Marijuanaut escapes earth to cultivate - Grow-room is church temple of the new stoner breed

Edited by sandman420 (12/16/21 05:34 AM)

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Re: Liquid culture [Re: Drboomer]
    #27584232 -

Yea, after what Notarealguy mentioned, I think I’m going to build a still air box and order some agar, more spore syringes, and LC jars. I’m just worried if the spawn bags will still be good, they’re still in original packaging and have been in a dark 50 degree room for about 2 weeks now.

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Re: Liquid culture [Re: sandman420]
    #27584236 -

I think it grew so fast because I injected 5ml in each jar then incubated at 82 degrees. I took some more pics today but it’s not too much different, although I noticed some growth on the side, I don’t know if that’s normal. I’m sure it will be more profound tomorrow so I will take new photos tomorrow and post the changes.

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Re: Liquid culture [Re: Quwo]
    #27584241 -

It'll probably take about 2 to 3 weeks to get to your second agar transfer, which is your third plate.    This is where you could drop the agar plate into some sterile water and blend it to inject, or make a master grain jar or whatever fits your situation best.

I prefer using whole agar plates to inoculate my spawn bags. They finish colonizing extremely quickly that way. Plus they don't throw off the moisture content of the grain like a LC can.


I'm not sure if the premade bags will hold up, I've never had any good luck with them.  They definitely have a shelf life, and you just never know if your getting old ones.


It's really best to prep everything yourself, so you know no corners have been cut and it's all fresh sub.

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Re: Liquid culture [Re: Notarealguy]
    #27584246 -

Yes I plan on making my own bags of spawn and substrate after I build an autoclave large enough to sterilize a bunch of bags at once. I’m using a 1:2 spawn to sub ratio with 10 pounds of grain and 20 pounds of horse manure per tub. I’m getting the bags from NorthSpore. Im only doing 10 tubs this time. My goal is to have 100 tubs going at once. Dial them in, after that scale up to 400 tubs staggered bi-weekly.

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Re: Liquid culture [Re: Quwo]
    #27584249 -

Hell yeah man, sounds like a plan.  One other thing I would recommend is looking into coco coir.  It's very easy to prep for bulk sub.  Pretty resistant to mold and bacteria.    It's Definitely easier Than manure, at least for me.

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Re: Liquid culture [Re: Quwo]
    #27584253 -

Ambitious goals.

Do the agar thing. If I can do it, you can do it.

You need anexic spawn. Clean and with limited genetics. You need agar work and cloning for this.  If your going to grow on such a large scale using MSS you will have heaps and heaps of smelly bacterial grains


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

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Re: Liquid culture [Re: Notarealguy]
    #27584271 -

I’m interested in hearing more about coco coir and casing I’m not experienced with using it. I was hoping the 1:2 ratio would colonize quickly so mold or bacteria should be at a minimum.

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Re: Liquid culture [Re: MysticMycologist]
    #27584277 -

Yea I don’t plan on using spore syringes after the first time. I don’t like the idea of them being mixed genetics anyways, I want to be able to dial it in on the agar. I’ve just been procrastinating on agar because I was under the impression I needed a laminar flow hood, but now I’m leaning more towards just using a still air box.

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