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OfflineArmillaria
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Registered: 03/16/15
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Last seen: 7 years, 4 months
Instant liquid culture
    #21415415 - 03/16/15 11:06 AM (9 years, 2 months ago)

Haven't seen this elsewhere so hope this helps.

I've found making liquid cultures a bit of a pain and they don't seem to have the quantity of mycelium in them that I see on an agar plate.

This method is quick to do and I use it for grain inoculations (also some bulk).

First with a large jar (say 500ml) pour some agar solution into the bottom (just 5mm or so). When it's solidified drop a piece of agar from a culture onto the new agar. Let it grow out (right-hand jar in the picture Hericium erinaceus).

When you need to do the liquid culture inoculation, pressure cook 100-200mls of nutrient solution (could use plain water if you want) with a mixer ball in the jar (left-hand jar).

After the mixer ball solution has cooled down pour the ball and the solution into the agar jar. Screw the lid down and shake like mad. The mixer ball chops up the agar into a mycelium soup.

Pour onto bulk substrate, grain or suck into a syringe.

The mixer balls are sold to body builders to mix protein powders into drinks. In the UK they cost about £2. I got mine from Amazon but they're on eBay and sports shops. They're just a spherical spiral of stainless steel.



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OfflineTonyHxC
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Re: Instant liquid culture [Re: Armillaria]
    #21415425 - 03/16/15 11:08 AM (9 years, 2 months ago)

:takingnotes:

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InvisibleGrey
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Re: Instant liquid culture [Re: Armillaria]
    #21415429 - 03/16/15 11:09 AM (9 years, 2 months ago)

you don't sterilize your agar first?


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:takingnotes:  AMU Q&A  :takingnotes:


If you don't have a plan of your own, you'll become a part of somebody else's.

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Offlinesilverstem
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Re: Instant liquid culture [Re: Armillaria]
    #21415436 - 03/16/15 11:12 AM (9 years, 2 months ago)

Yes its like glc.. Except alc. Nice but I don't like lcs but still nice.


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Shroomery needs a gun forum!!!!!!!!! CAN WE HAVE ONE?????

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OfflineTonyHxC
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Re: Instant liquid culture [Re: Grey]
    #21415442 - 03/16/15 11:15 AM (9 years, 2 months ago)

Quote:

Grey said:
you don't sterilize your agar first?





I assumed he was sterilizing the agar when ever he made it in the jar, maybe I am wrong though.. I really hope he is sterilizing it.

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OfflineBlunderSoftly
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Re: Instant liquid culture [Re: Grey]
    #21415465 - 03/16/15 11:22 AM (9 years, 2 months ago)

He's describing a method of making liquid culture from agar already colonized, not making an agar culture.

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OfflineArmillaria
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Re: Instant liquid culture [Re: silverstem]
    #21415531 - 03/16/15 11:39 AM (9 years, 2 months ago)

Yes the agar culture is sterilised.
1. Pour nutrient solution into bottom of right-hand jar. Add agar powder.
2. Sterilise in PC, cool, set.
3. Inoculate
Wait (few days) till grown out (this is now the right-hand jar in picture).
4. Sterilise mixer ball in nutrient solution and let cool (left hand jar).
Add left jar to right jar, shake, etc.

You could do both PC sterilisations at the same time and just leave the mixer ball jar till you need it (when the mycelium's grown out on the agar).

PS left jar is liquid, no agar in it.

Edited by Armillaria (03/16/15 11:45 AM)

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Offlineblackout
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Re: Instant liquid culture [Re: Armillaria]
    #21416582 - 03/16/15 03:51 PM (9 years, 2 months ago)

Quote:

Armillaria said:pressure cook 100-200mls of nutrient solution (could use plain water if you want) 



most would use water. People want to minimize the amount of nutrients going into their innoculating liquids, sugars and liquid invite contams. Unless you were planning to grow out more myc in the LC.

You could minimize your agar volume for this reason too, or at least spread it out more. If agar is thick it can be rolled around a jar when its just about to set so it has far more surface area to grow on. I am using these sharwood square jars as petri dishes. When tiled to one side you get a lot of surface area, and can look down from above and see growth clearly, with no distortion due to looking into rounded glass.


I have seen those mixer balls and thought they would be too light and blunt to cut up the myc well, esp a mat of myc on agar. Have you tried other things? how does it compare? I have used stones, shards of glass, heavy stainless ball bearings. The ball bearing is too blunt to cut but can work well with the glass.

You could add your wire ball to the agar jar from the start. I expect it might be a bit haphazard trying to pour the liquid with the wire ball into the agar jar, i.e. I expect the ball just suddenly flies out of the jar.

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OfflineArmillaria
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Re: Instant liquid culture [Re: blackout]
    #21416700 - 03/16/15 04:17 PM (9 years, 2 months ago)

Excellent idea - spreading agar round the inside of the jar. I'll try that next time.

I thought of steel ball bearings but they could break the glass. The mixer balls are light but seem to cut 1% agar well. I've only used mixer balls (nothing else). They're cheap so it's worth getting one to experiment with - nullius in verba.

I use nutrient solution rather than water as I normally go on to pour the soup onto grain for spawn. I get fast growth and no contamination.

I don't put the mixer ball in the agar jar as it means I can't use it while the mycelium grows out (several days). If I sterilise it with the non-agar solution it's only in use for a few minutes so I don't need to buy lots of them.

The Sharwoods jar's necks are a bit small for the mixer ball (ball is 5cm across). I use mayo jars.

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OfflineTonyHxC
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Re: Instant liquid culture [Re: Armillaria]
    #21416866 - 03/16/15 04:51 PM (9 years, 2 months ago)

When you say Nutrient Solution do you mean like Water and Honey?

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OfflineArmillaria
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Re: Instant liquid culture [Re: TonyHxC]
    #21418944 - 03/17/15 01:47 AM (9 years, 2 months ago)

No. All the liquids (for shaking and for agar) are 1% malt extract (from beer making kit), 1% yeast extract (UK Marmite - like vegemite), 10% vinegar and 0.2% alanine.

I've found 10% vinegar seems to prevent contamination. I'm a (used to be) postgrad. level biochemist/chemist and and I'm inoculating in a pretty chaotic kitchen and I've had almost no contamination. I'm talking 1 or 2 percent of over 100 inoculations over 3 months.

This is for Hericium erinaceus so vinegar may screw other species. I think I can get the level of vinegar down but still get the protective affect.

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