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Yuggoth
Mi-Go Cultivator



Registered: 03/04/23
Posts: 493
Loc: Lost Carcosa
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Re: SEMI-SOLID LIQUID CULTURE [Re: altford78] 1
#28344394 - 06/02/23 12:08 PM (7 months, 23 days ago) |
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How much spinning you need depends on what the mechanism of the benefit is, I guess.
For example when you are growing yeast for beer, they need oxygen to replicate. Spinning the culture increases the amount of oxygen that is dissolved into the solution.
But yeast need that oxygen mostly for creating the compounds that make up their cell walls. Mycelium may not be the same.
Well, let's look that up... OK after some googling, it looks like all fungi synthesize the same oxygen-containing compound for their cell walls. It's called ergosterol. (Other sterols are surely in the mix too, ergosterol is just dominant.)
It seems reasonable that mycelium, which also synthesize ergosterol, would benefit from dissolved oxygen the same way that yeast do.
So how much do you have to spin the culture to keep the optimum level of oxygen dissolved? Beats me, but there is probably enough data out there to figure it out.
But if you only do it 5 minutes a day, you may be undershooting the oxygen level for optimum growth... Whereas if you do it continuously, you are more likely to supply enough. Maybe you'll provide more than enough, but that seems harmless.
There are a lot of other variables... Temperature, surface area, permeability of whatever filter the LC jar uses, myc species...
But from what I know, if I am gonna spin it, I am gonna spin it gently but constantly. If I have more than one LC jar cooking, I let them take turns, switching jars every 8-12 hours when I walk by.
YMMV.
-------------------- We have not succeeded in answering all our problems. The answers we have found only serve to raise a whole set of new questions. In some ways we feel we are as confused as ever, but we believe we are confused on a higher level and about more important things. -- Earl C. Kelley Things I really wish I knew when I started // Vacuum sealer discussion thread // Shroomery gif zoo
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altford78
What do I put here

Registered: 05/09/23
Posts: 890
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Re: SEMI-SOLID LIQUID CULTURE [Re: Yuggoth]
#28344400 - 06/02/23 12:19 PM (7 months, 23 days ago) |
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Interesting, I will look into this, thanks!
Just so there is no misunderstanding, you have your LC spinning on 12h on/12h off cycles?
When you say "gently", how do you quantify gently?
-------------------- Mold cultivator extraordinaire I also dabble in bacteria I'm also a retard sometimes
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Yuggoth
Mi-Go Cultivator



Registered: 03/04/23
Posts: 493
Loc: Lost Carcosa
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Re: SEMI-SOLID LIQUID CULTURE [Re: altford78]
#28344422 - 06/02/23 12:46 PM (7 months, 23 days ago) |
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I stir constantly but since I only have one stir plate, if I am doing more than one jar they take turns.
"Gently" means the vortex at the top is no more than about 1/2" deep. I never stir to the point that bubbles are introduced but everything is moving around freely. This is about 20-25% power on my Amazon stir plate.
Honestly I think you could get the same oxygen dissolving benefits with minimal surface motion. Look at a big aquarium, where the ratio of surface area to volume is worse... They stay oxygenated with relatively little surface motion. But stronger stirring keeps the myc from clumping up, too.
-------------------- We have not succeeded in answering all our problems. The answers we have found only serve to raise a whole set of new questions. In some ways we feel we are as confused as ever, but we believe we are confused on a higher level and about more important things. -- Earl C. Kelley Things I really wish I knew when I started // Vacuum sealer discussion thread // Shroomery gif zoo
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altford78
What do I put here

Registered: 05/09/23
Posts: 890
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Re: SEMI-SOLID LIQUID CULTURE [Re: Yuggoth]
#28344438 - 06/02/23 12:59 PM (7 months, 23 days ago) |
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Cool, thanks. I've spun mine at max rpm a couple times and yeah a lot of bubbles but the myc seems to recover pretty well.
-------------------- Mold cultivator extraordinaire I also dabble in bacteria I'm also a retard sometimes
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Yuggoth
Mi-Go Cultivator



Registered: 03/04/23
Posts: 493
Loc: Lost Carcosa
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Re: SEMI-SOLID LIQUID CULTURE [Re: altford78] 1
#28344445 - 06/02/23 01:06 PM (7 months, 23 days ago) |
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If I have a culture sitting off the plate and it clumps up, I give it a good hard spin to break it up before I try drawing it into a syringe. I agree the bubbles don't hurt in normal stirring... I just don't want to look at them, and you might hear the stir bar rattle at higher speeds too.
-------------------- We have not succeeded in answering all our problems. The answers we have found only serve to raise a whole set of new questions. In some ways we feel we are as confused as ever, but we believe we are confused on a higher level and about more important things. -- Earl C. Kelley Things I really wish I knew when I started // Vacuum sealer discussion thread // Shroomery gif zoo
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altford78
What do I put here

Registered: 05/09/23
Posts: 890
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Re: SEMI-SOLID LIQUID CULTURE [Re: Yuggoth]
#28346938 - 06/04/23 10:57 AM (7 months, 21 days ago) |
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are there any visual cues to indicate that the LC is ready to go to grain? I mean, at what point are you satisfied with the myc growth in the bottle?
also, if you want to test it, do you aspirate with a syringe or free pour as little as possible on an agar plate? I'm using unmodified gl45 lids so I'm just thinking of pouring on a plate.
-------------------- Mold cultivator extraordinaire I also dabble in bacteria I'm also a retard sometimes
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Yuggoth
Mi-Go Cultivator



Registered: 03/04/23
Posts: 493
Loc: Lost Carcosa
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Re: SEMI-SOLID LIQUID CULTURE [Re: altford78]
#28346951 - 06/04/23 11:11 AM (7 months, 21 days ago) |
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To test I extract with a syringe. I did that through the SHIP (and it was the only use of the SHIP post-sterilization). But, I have also taken off the lid to get a needle into the LC and that worked OK too.
I cannot imagine freely pouring the tiny amount needed.
-------------------- We have not succeeded in answering all our problems. The answers we have found only serve to raise a whole set of new questions. In some ways we feel we are as confused as ever, but we believe we are confused on a higher level and about more important things. -- Earl C. Kelley Things I really wish I knew when I started // Vacuum sealer discussion thread // Shroomery gif zoo
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altford78
What do I put here

Registered: 05/09/23
Posts: 890
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Re: SEMI-SOLID LIQUID CULTURE [Re: Yuggoth]
#28346964 - 06/04/23 11:27 AM (7 months, 21 days ago) |
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cool, thanks. Do you cool the needle in the LC ?
Yeah I was also wondering if it's possible to pour that little into a petri. will probably make a mess.
what needle gauge are you using? 14-16g should be fine I think, I have some long blunt nose 14g that should do nicely.
-------------------- Mold cultivator extraordinaire I also dabble in bacteria I'm also a retard sometimes
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Yuggoth
Mi-Go Cultivator



Registered: 03/04/23
Posts: 493
Loc: Lost Carcosa
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Re: SEMI-SOLID LIQUID CULTURE [Re: altford78]
#28346978 - 06/04/23 11:38 AM (7 months, 21 days ago) |
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At the time I did this I used a new sterile 16 ga needle, so I did not have to heat it. 16 ga was bigger than needed and I didn't have anything smaller at the time, but it worked OK. Today I would use a 21 ga instead just to minimize wear and tear on the SHIP... Those are the two sizes I currently stock.
(I never flame-sterilize a needle if the needle needs to go into a SHIP or solution, I will just put on a new one. They are dirt cheap and that way I don't have to even think about cooling the needle. The heat also makes them less sharp, which sucks. If I am dispensing solution from the syringe without needing the syringe to touch another object, like dripping LC onto a plate, then I will torch it and cool it by squirting out extra solution. YMMV.)
-------------------- We have not succeeded in answering all our problems. The answers we have found only serve to raise a whole set of new questions. In some ways we feel we are as confused as ever, but we believe we are confused on a higher level and about more important things. -- Earl C. Kelley Things I really wish I knew when I started // Vacuum sealer discussion thread // Shroomery gif zoo
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A.k.a
Stranger



Registered: 10/27/19
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Re: SEMI-SOLID LIQUID CULTURE [Re: Yuggoth]
#28347158 - 06/04/23 02:12 PM (7 months, 21 days ago) |
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Ime by five days it’s worth using. Definitely not as thick as it will be in another five days but I usually get fast growth even when it doesn’t look super colonized.
What I’ve been doing is using a ship and filling a 20 or 30ml syringe, dripping some on a plate, then capping the syringe up and once the plate shows clean using the syringe. Free pour works well too though.
--------------------
LAGM2020     
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Yuggoth
Mi-Go Cultivator



Registered: 03/04/23
Posts: 493
Loc: Lost Carcosa
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Re: SEMI-SOLID LIQUID CULTURE [Re: A.k.a]
#28347181 - 06/04/23 02:35 PM (7 months, 21 days ago) |
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Quote:
A.k.a said: What I’ve been doing is using a ship and filling a 20 or 30ml syringe, dripping some on a plate, then capping the syringe up and once the plate shows clean using the syringe.
Good thinkin'!
-------------------- We have not succeeded in answering all our problems. The answers we have found only serve to raise a whole set of new questions. In some ways we feel we are as confused as ever, but we believe we are confused on a higher level and about more important things. -- Earl C. Kelley Things I really wish I knew when I started // Vacuum sealer discussion thread // Shroomery gif zoo
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altford78
What do I put here

Registered: 05/09/23
Posts: 890
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Re: SEMI-SOLID LIQUID CULTURE [Re: Yuggoth]
#28350358 - 06/07/23 02:46 AM (7 months, 18 days ago) |
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Since the agar makes the LC turbid by itself, what kind of visual indicators do you use for bacterial contamination? The old "put something with letters behind the bottle and if you can't read it, it's bacterial" doesn't really work here.
-------------------- Mold cultivator extraordinaire I also dabble in bacteria I'm also a retard sometimes
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Smoothcat
Renegade-master



Registered: 10/07/15
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Re: SEMI-SOLID LIQUID CULTURE [Re: altford78]
#28361420 - 06/15/23 07:52 PM (7 months, 10 days ago) |
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Made 3 of these last night - it was late and accidentally put 4g lme and 4g agar in each jar  Ended up making Super Solid Liquid Culture

Anyway I put all that in one jar and in the fridge and will try and dilute that at a later stage. Made 3 more with .4g lme and .4g agar
Got a couple of cubes and one pan
The wedges look like little jelly fish suspended in the soup - I like it
Be interested to see how this goes
-------------------- Back once again with the ill behaviour Links I like
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