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superfunkibey
Stranger

Registered: 11/29/10
Posts: 141
Loc: Philippines
Last seen: 8 years, 10 months
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Liquid Culture Question
#14227961 - 04/02/11 07:43 PM (12 years, 10 months ago) |
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I have read some of the LC teks in this site and was wondering why there is a need to incubate/grow them in the honey mixture?
I read in Stamet's book that he just blenders the pda mat, adds it to sterile water for the proper dilution afterwards he injects this straight into the substrate.
I would appreciate being enlightened about this.
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Terry M
Stranger in a Strange Land



Registered: 06/18/10
Posts: 1,502
Loc: Rhode Island
Last seen: 9 years, 1 month
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Quote:
superfunkibey said: I read in Stamet's book that he just blenders the pda mat, adds it to sterile water for the proper dilution afterwards he injects this straight into the substrate.
Are you sure that you saw this? Stamets' GGMM goes into a fair amount of LC detail. He talks about using nutrient-enriched water, with malt sugar and other things.
-------------------- Liberté, égalité, humidité.
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fungus_tao
Hah Zah!



Registered: 05/10/06
Posts: 1,856
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Re: Liquid Culture Question [Re: Terry M]
#14230336 - 04/03/11 09:19 AM (12 years, 10 months ago) |
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The first time I ever made an LC was with Elm oyster culture. I completely skipped the incubation step. I used nutrient water, but just blended an agar wedge in a mason jar and immediately filled a syringe and injected into jars. All jars colonized without contams. They were very very slow to get started. But that would suggest that you could use plain water and blend an agar wedge into it dispersing the mycellium on the wedge into the water and then using that. Of course you'll get better results if you use nutrient solution and incubate. Hit the grain running vs. long recovery period.
-------------------- Follow the light The Light is your guide.
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superfunkibey
Stranger

Registered: 11/29/10
Posts: 141
Loc: Philippines
Last seen: 8 years, 10 months
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Re: Liquid Culture Question [Re: Terry M]
#14289067 - 04/14/11 09:39 AM (12 years, 10 months ago) |
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Quote:
Terry M said:
Quote:
superfunkibey said: I read in Stamet's book that he just blenders the pda mat, adds it to sterile water for the proper dilution afterwards he injects this straight into the substrate.
Are you sure that you saw this? Stamets' GGMM goes into a fair amount of LC detail. He talks about using nutrient-enriched water, with malt sugar and other things.
Well, I read it in the Mushroom Cultivator, though maybe there has been improvements.
Kinda uncertain which LC method would be best to try though
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Terry M
Stranger in a Strange Land



Registered: 06/18/10
Posts: 1,502
Loc: Rhode Island
Last seen: 9 years, 1 month
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Quote:
superfunkibey said:
Quote:
Terry M said:
Quote:
superfunkibey said: I read in Stamet's book that he just blenders the pda mat, adds it to sterile water for the proper dilution afterwards he injects this straight into the substrate.
Are you sure that you saw this? Stamets' GGMM goes into a fair amount of LC detail. He talks about using nutrient-enriched water, with malt sugar and other things.
Well, I read it in the Mushroom Cultivator, though maybe there has been improvements.
Kinda uncertain which LC method would be best to try though
Yup, right you are! In TMC, Stamets actually writes about two techniques. The first uses the mycelial mat on top of a colonized grain jar. In the second, he blends a colonized agar plate. Both use just sterilized water.
I think a lot of folks around here start their LCs from a 10 ml spore or culture syringe. That's what I did. For this method, I believe you need the nutrients to greatly expand the mycelium. When you first inject with the syringe, you don't see anything going on in the jar. But after a few days, you get rich clouds of fluffy mycelium floating in the liquid.
Personally, I just ditched my refrigerated LC jars now that I've gotten comfortable with agar. Big LC jars provide an opportunity to loose your entire culture to contamination without even knowing it. I've already experienced this. Plus they don't really store for very long. I am right now switching to GLC in syringes for medium-term refrigerated storage (thanks to Doc_T and RogerRabbit!) and distilled water long-term storage (thanks to FreeSporePrints!).
-------------------- Liberté, égalité, humidité.
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ife
Stranger Danger


Registered: 04/10/11
Posts: 116
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Re: Liquid Culture Question [Re: Terry M]
#14289377 - 04/14/11 11:00 AM (12 years, 10 months ago) |
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Oh All Natural Molasses works well for LC. About 1 tablespoon ish mixed in water in a 1 qt jar with a self-healing syringe hole in the lid. PC'd for 1 hour. Inject with spore syringe once the jars have cooled to room temp. Wait 2 weeks. Use. Or you could wait a bout 3 days and use. Though waiting the 2 weeks allows the myc to gain more strength and speed in colonization. No contams so far with the molasses method. Maybe I am just lucky so far.
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mycofanatic



Registered: 06/12/08
Posts: 322
Last seen: 10 years, 12 days
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Re: Liquid Culture Question [Re: ife]
#14300238 - 04/16/11 10:02 AM (12 years, 10 months ago) |
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Many of these methods will work. I like to make malt extract LC's and inject them with 10-20 ml of a mature LC (from vacutainers) through injector ports. I prefer this because there's only one opportunity for contaminants to get in, and a very low likely hood if you have good aseptic technique.
You start running into problems when you're taking the lid off 2-3 times, unless you have a hepa and a sterile working environment. Because its much more difficult to detect contams in LC's compared to agar plates.
Try them all and see what works best for you.
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PitcherCrab
Crescent Fresh



Registered: 12/02/06
Posts: 2,446
Loc: The bottom of the sea.
Last seen: 1 year, 2 months
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Re: Liquid Culture Question [Re: mycofanatic]
#14302927 - 04/16/11 07:40 PM (12 years, 10 months ago) |
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-------------------- PC's LAGM 2021 (TOC & TWC)P. natalensis Growlog 2021Pans for PC Fall 2021 Growlog “I must not fear. Fear is the mind-killer. Fear is the little death that brings total obliteration. I will face my fear. I will permit it to pass over me and through me. And when it has gone past I will turn the inner eye to see its path. Where the fear has gone there will be nothing. Only I will remain.” - Bene Gesserit Litany Against Fear
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Humility
Working on it



Registered: 10/07/08
Posts: 6,745
Last seen: 7 years, 10 days
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Re: Liquid Culture Question [Re: PitcherCrab]
#14304917 - 04/17/11 07:05 AM (12 years, 10 months ago) |
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