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Shroomburger
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Liquid media questions
#18868837 - 09/20/13 05:49 PM (10 years, 5 months ago) |
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Hi All,
I want to grow Hericium coralloides and H. americanum in liquid nutrient media. One study says H. erinaceus was grown on "SGC" which apparently is glucose, starch, corn steep liquor KH2PO4, CuSO4, ZnCl2, thiamine chloride and water, but they don't say the proportions. Does anyone know the proportions for this mix? Or perhaps a better, or suitable alternative? Just anything that is good for mushrooms in general (going in blind).
SB
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liamtheloser
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Registered: 06/07/06
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You are just wanting to make a liquid culture for inoculating sawdust bags or something?
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Shroomburger
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Actually, I want to grow the mycelium primarily to extract from for a research project. But also, as a side project, I want to grow them on sawdust, yes.
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liamtheloser
Advanced Idiot

Registered: 06/07/06
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Have you tried standard solutions like Karo corn syrup?
I find a good media for growing mycelium is 1 tablespoon of Karo light corn syrup and 1 teaspoon of instant coffee per half pint jar of water.
If you wanted to get fancy you could use light malt or potato dextrose or anything you'd use in agar, but instead use in water (sans agar)
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Shroomburger
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I haven't tried anything yet. I just got one of my two spp. into culture on agar and I would like to transfer it to liquid media soon. Really?? Just corn syrup and coffee? Ya have good success with that eh? With Hericium by chance? That sounds simple 'nuff, but I am willing to go further than that if it will increase my chance of success. Would you suggest adjusting pH or adding additional supplements? I can get maltose and potato dextrose at my school (Proportions?). What else?
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liamtheloser
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Registered: 06/07/06
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http://www.shroomery.org/forums/showflat.php/Number/12826752
Yes, if it works on agar, it will work in the liquid. I haven't done hericium yet, but will be soon with lions mane. It has worked with everything else I've tried though!
I don't have easy access to malt dextrose or potato dextrose so I use what I have, which is light corn syrup and powdered coffee 
You should see my agar mix, it's corn syrup and bakers yeast boiled into the agar.
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drake89
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Registered: 06/26/11
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i just tried and failed with the recipe in ggmm which is something like malt extract, yeast extract, water, and a bit of substrate plus blended agar dishes. That's not to say it doesn't work, I just haven't had much luck since my fist attempt which was basically sugar water as mentioned above.
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OICU812
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Registered: 11/06/11
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Quote:
Shroomburger said: I haven't tried anything yet. I just got one of my two spp. into culture on agar and I would like to transfer it to liquid media soon. Really?? Just corn syrup and coffee? Ya have good success with that eh? With Hericium by chance? That sounds simple 'nuff, but I am willing to go further than that if it will increase my chance of success. Would you suggest adjusting pH or adding additional supplements? I can get maltose and potato dextrose at my school (Proportions?). What else?
A Stir plate would greatly assist in the propagation of the liquid-grown mycelium. I am a long time home brewer and making a liquid starter is very much like making liquid mycelium media. Here is a very good paper by Dr. Maribeth Raines, Ph.D.: "Yeast Propagation and Maintenance: Principles and Practices"
-------------------- -------------- "Democracy is two wolves and a lamb voting on what to have for lunch. Liberty is a well-armed lamb contesting the vote!" --Benjamin Franklin "Those who give up liberty for security won't have, or deserve, either.". . . Benjamin Franklin ----> Read: The Fight of our Lives - Defeating the Ideological War Against the West - by Victor Davis Hanson
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Forrester
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Re: Liquid media questions [Re: OICU812]
#18869455 - 09/20/13 08:28 PM (10 years, 5 months ago) |
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Quote:
OICU812 said: A Stir plate would greatly assist in the propagation of the liquid-grown mycelium. I am a long time home brewer and making a liquid starter is very much like making liquid mycelium media. Here is a very good paper by Dr. Maribeth Raines, Ph.D.: "Yeast Propagation and Maintenance: Principles and Practices"
Any tips on LC making, without a stir bar? I just don't have one. Inoculation port and some gas exchange, and shake when I can, will that work? All I've got are filter discs, will they work if I swirl and be careful not to get wet? Sorry not trying to thread-jack but it seems on topic
-------------------- Repugnant is a creature who would squander the ability to lift an eye to heaven, conscious of his fleeting time here. ------------------- Have some medicinal mushrooms and want to get the most out of them? Try this double extraction method.
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OICU812
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Re: Liquid media questions [Re: Forrester]
#18869699 - 09/20/13 09:16 PM (10 years, 5 months ago) |
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Quote:
Forrester said:
Any tips on LC making, without a stir bar? I just don't have one. Inoculation port and some gas exchange, and shake when I can, will that work? All I've got are filter discs, will they work if I swirl and be careful not to get wet? Sorry not trying to thread-jack but it seems on topic 
Sure. Stir plates are not mandatory, they just greatly increase the mycelium count.
Here are a few tips:
- Sterilize everything but the mycelium. LC is very unforgiving of any contamination.
- Use a good jar lid. Here is a good Tek on making them.
- Use only SFDs -or- Whatmans for air exchange.
- I like injection ports over homemade silicone ports.
- TL's Simple Liquid Culture (LC) Tek is a pretty good Tek with solid procedures.
That's about it. Not complicated. Most important thing is being anal about sterility and cleanliness.
-------------------- -------------- "Democracy is two wolves and a lamb voting on what to have for lunch. Liberty is a well-armed lamb contesting the vote!" --Benjamin Franklin "Those who give up liberty for security won't have, or deserve, either.". . . Benjamin Franklin ----> Read: The Fight of our Lives - Defeating the Ideological War Against the West - by Victor Davis Hanson
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liamtheloser
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Re: Liquid media questions [Re: OICU812]
#18869764 - 09/20/13 09:35 PM (10 years, 5 months ago) |
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You don't have it sitting on the stir plate 24/7, do you? All I use my stir plate for is to break it up once ina while to help aspread, or to break it up before I fill a syringe.
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Forrester
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Re: Liquid media questions [Re: OICU812]
#18870940 - 09/21/13 06:22 AM (10 years, 5 months ago) |
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Thanks OICU!
-------------------- Repugnant is a creature who would squander the ability to lift an eye to heaven, conscious of his fleeting time here. ------------------- Have some medicinal mushrooms and want to get the most out of them? Try this double extraction method.
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t3chnobily
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I keep my working masters on LC in quart jars with plastic lids with rubber rings, GE and injections ports. I use a teaspoon of MEA agar mix to 2/3 of a quart of hot water. This gives me a viscous liquid that holds air bubbles when I give them their daily swirl.
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OICU812
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Registered: 11/06/11
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Quote:
liamtheloser said: You don't have it sitting on the stir plate 24/7, do you? All I use my stir plate for is to break it up once ina while to help aspread, or to break it up before I fill a syringe.
Actually yes, I do stir 24/7. It makes an exponential difference in the cell count. The purpose of using a stir plate is to increase the saturation of oxygen in the liquid. Yeast and mycelium are very close cousins. They both consume oxygen and give off carbon dioxide and water. Figure 1 in Dr. Raines article depicts the effect of airlock (not stirred or shaken) -vs- shaken -vs- aerated -vs- stirred. In the description under Figure 1, she describes "continuously stirred on a magnetic stir plate".
The vortex created by the stir bar draws air down into the liquid and creates pressure against the vortex wall aiding in the absorption of oxygen. Some of my fellow brewers are so anal about this that they have small oxygen tanks and give the starter (and the wort) a short blast of pure oxygen when they first pitch the yeast.
-------------------- -------------- "Democracy is two wolves and a lamb voting on what to have for lunch. Liberty is a well-armed lamb contesting the vote!" --Benjamin Franklin "Those who give up liberty for security won't have, or deserve, either.". . . Benjamin Franklin ----> Read: The Fight of our Lives - Defeating the Ideological War Against the West - by Victor Davis Hanson
Edited by OICU812 (09/21/13 06:52 AM)
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liamtheloser
Advanced Idiot

Registered: 06/07/06
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Re: Liquid media questions [Re: OICU812]
#18872045 - 09/21/13 12:57 PM (10 years, 5 months ago) |
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So the stirring doesn't hurt the mycelium? I figured it would need time between stirs to recover, but if not I'll keep mine on the stir plate all day.
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OICU812
NC Tree Farm owner


Registered: 11/06/11
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Quote:
liamtheloser said: So the stirring doesn't hurt the mycelium? I figured it would need time between stirs to recover, but if not I'll keep mine on the stir plate all day.
Doesn't hurt anything. Stir continuously.
-------------------- -------------- "Democracy is two wolves and a lamb voting on what to have for lunch. Liberty is a well-armed lamb contesting the vote!" --Benjamin Franklin "Those who give up liberty for security won't have, or deserve, either.". . . Benjamin Franklin ----> Read: The Fight of our Lives - Defeating the Ideological War Against the West - by Victor Davis Hanson
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drake89
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Re: Liquid media questions [Re: OICU812]
#18872745 - 09/21/13 03:48 PM (10 years, 5 months ago) |
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everyone asking questions in this thread should buy a copy of Growing gourmet and medicinal mushrooms by paul stamets. this topic is one of the early ones covered in the book. he's all about eberbach (sp?) blenders, which are just $750 stainless steel blenders. You can substitute this for an oster blender with the blade attached to a small mouth mason jar. Use the regular metal jar ring instead of the plastic oster base, as it will melt in the PC.
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Shroomburger
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Re: Liquid media questions [Re: OICU812]
#18872812 - 09/21/13 04:06 PM (10 years, 5 months ago) |
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Stir plates I have plenty of! At what frequency should it stir??
Edited by Shroomburger (09/21/13 04:24 PM)
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OICU812
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Re: Liquid media questions [Re: drake89]
#18873236 - 09/21/13 06:06 PM (10 years, 5 months ago) |
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Quote:
drake89 said: everyone asking questions in this thread should buy a copy of Growing gourmet and medicinal mushrooms by paul stamets. this topic is one of the early ones covered in the book. he's all about eberbach (sp?) blenders, which are just $750 stainless steel blenders. You can substitute this for an oster blender with the blade attached to a small mouth mason jar. Use the regular metal jar ring instead of the plastic oster base, as it will melt in the PC.
Thanks, Drake89. Yes, GGMM Pgs. 137-139 detail the liquid culture process. I assumed everyone was familiar with getting the mycelium ready for the stir plate and was only addressing the benefits of it's use. Actually the Eberbach or blender is used to chop mycelium from a petri dish to get it ready to be placed in an Erlenmeyer for stirring.
Quote:
Shroomburger said: Stir plates I have plenty of! At what frequency should it stir??
Stamets, in the text mentioned above, says 100-200 RPM but I just adjust until I get a good looking vortex.
-------------------- -------------- "Democracy is two wolves and a lamb voting on what to have for lunch. Liberty is a well-armed lamb contesting the vote!" --Benjamin Franklin "Those who give up liberty for security won't have, or deserve, either.". . . Benjamin Franklin ----> Read: The Fight of our Lives - Defeating the Ideological War Against the West - by Victor Davis Hanson
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drake89
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Re: Liquid media questions [Re: OICU812]
#18873305 - 09/21/13 06:32 PM (10 years, 5 months ago) |
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Quote:
OICU812 said:
Quote:
drake89 said: everyone asking questions in this thread should buy a copy of Growing gourmet and medicinal mushrooms by paul stamets. this topic is one of the early ones covered in the book. he's all about eberbach (sp?) blenders, which are just $750 stainless steel blenders. You can substitute this for an oster blender with the blade attached to a small mouth mason jar. Use the regular metal jar ring instead of the plastic oster base, as it will melt in the PC.
Thanks, Drake89. Yes, GGMM Pgs. 137-139 detail the liquid culture process. I assumed everyone was familiar with getting the mycelium ready for the stir plate and was only addressing the benefits of it's use. Actually the Eberbach or blender is used to chop mycelium from a petri dish to get it ready to be placed in an Erlenmeyer for stirring.
Quote:
Shroomburger said: Stir plates I have plenty of! At what frequency should it stir??
Stamets, in the text mentioned above, says 100-200 RPM but I just adjust until I get a good looking vortex.
How long do you incubate? i tried 48 hours and it didn't seem to accelerate anything. How do the 'pros' that sell syringes get their so clear? It's usually malt extract solution with a little 'worm' of more solid myc in it. Maybe they don't use all of the additives recomended in ggmm, or perhaps there's a sterile filter.
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