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OfflineDrShroomAholic
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Opions Please "Liquid Growth Medium Q/A"
    #1539955 - 05/11/03 11:15 PM (13 years, 6 months ago)

What is your opion ?

Mycelium can be cultured in a liquid (as opposed to a solid like vermiculite/brown rice flour) which you can then use to easily inoculate substrate and start mycelium growth much faster than if you were using spores ?

You should first go to your local homebrew supply store and buy the lightest colored powdered malt extract they have. Mix 4% (by weight) malt extract powder with distilled water (boil it, and then mix in the powder. No need to let it cool to room temperature first). Pour this medium into any large jar with a metal lid, and make one small hole in the lid with a nail. Cover the top tightly with aluminum foil, and either boil the jar in water for an hour (be sure to put the jar on a washcloth and not directly on the bottom of the pot) or use a pressure cooker set at 15psi for 20 minutes. Let the jar cool overnight.

The next day take your spore syringe, stick the needle through the hole in the lid, and squeeze in the solution. Cover the top of the jar with aluminum foil again, only not too tightly since you'll want to allow gas exchange. Keep the jar somewhere warm and dark, and agitate it every day. You should notice the solution becoming milky, and towards the end of the incubation you'll see fluffy pieces of mycelium floating around in it. At this point you can keep it in the fridge until you're ready to use it. When you want to start a mushroom culture somewhere, flame-sterilize the needle of a 10cc syringe, let it cool, stick it through the hole in the top of the lid, and pull in as much solution as you need. You can then inject this into your substrate and it will start growing immediately. Any time you run low on mycelium solution, go through the same process with another jar only instead of injecting spores inject the last of your mycelium solution. Be on the lookout for any contamination!

If this indeed true why waste time on the PF Tek and make cakes ? seems you could shorten your harvest time going this route ? Or am i missing somthing ?

Please post any good or bad experinces and your opion's

This was copy'd out of the FAQ this is not my tek I am just using it as a examble. :grin:


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Open wide and shut your eye's stick out your toung you are in for a suprise chew a cap and chew a stem empty your mouth say AMEN and sit on down and let the fun begin your about to leave reality time and time again.


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Invisiblemicro
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Re: Opions Please [Re: DrShroomAholic]
    #1540025 - 05/11/03 11:36 PM (13 years, 6 months ago)

Sounds like it should work if you are completely sterile, but if it isn't clear like dextrose water it will be harder to distinguish contaminants. Just make sure to leave some room for air and shake a bit here and there to get the air to dissolve.

The Difco Manual has this recipe for malt extract broth:

Per liter:

Malt extract 6g
Maltose 1.8g
Dextrose 6g
Yeast Extract 1.2g

Ster @ 121C/250F (~15 psi) for 15 min

They pH it to 4.7 to be selective for fungi.
________________

Don't see why just malt extract wouldn't work, though.... Just keep it sterile -- maybe use a septum plug or something....

--
Micro


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InvisibleSixTango
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Re: Opions Please "Liquid Growth Medium Q/A" [Re: DrShroomAholic]
    #1540283 - 05/12/03 12:55 AM (13 years, 6 months ago)

My foaf has used a light dry malt gold along the lines to state. One added step was to heat it in water, so it disolves, then strain in through coffee filters into a jar, then PC. However, the liquid culture was NOT injected into any substrate. It was used to to start WBS jars and/or bags of spawn. Never tried it straight into a substrate.
6T :wink:
 


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Offlinedebianlinux
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Re: Opions Please [Re: SixTango]
    #1541058 - 05/12/03 08:15 AM (13 years, 6 months ago)

I've shot plenty of BRF jars with LC. Practically all my work involves LC as a starter. The main advantage is that you fo get about a 2-3 day head start than you would with spores. Like 6T said, it is especially effective in grains primarily because of the way it initially distributes itself when you shake it, not something that can be done with BRF.


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OfflineDrShroomAholic
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Re: Opions Please [Re: debianlinux]
    #1541146 - 05/12/03 10:17 AM (13 years, 6 months ago)

How about popcorn,, What casing mix would you use in LC ?


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Open wide and shut your eye's stick out your toung you are in for a suprise chew a cap and chew a stem empty your mouth say AMEN and sit on down and let the fun begin your about to leave reality time and time again.


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OfflineStopThat
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Re: Opions Please [Re: DrShroomAholic]
    #1541557 - 05/12/03 01:38 PM (13 years, 6 months ago)

It's great on popcorn. Once you get the grain colonized,you can case with any of the ones listed in the casing FAQ.Coco coir/Verm is a good one,it's simple.


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Invisiblepussycontrol
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Re: Opions Please [Re: StopThat]
    #1542525 - 05/12/03 07:33 PM (13 years, 6 months ago)

afoaf. Hes made it a few times. its actually very easy to do. ive heard it cuts time of colonization by like 2 weeks ahead, ill update that tonigh, afoaf just used it a week ago


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OfflineShaMan1988
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Re: Opions Please "Liquid Growth Medium Q/A" [Re: DrShroomAholic]
    #1546390 - 05/13/03 09:55 PM (13 years, 6 months ago)

hey.
no offence. but you wrote what you said in a way as though you just invented that tek. a good tek to follow would be one written by no other than By Nan (Nanook) on Friday, November 02, 2001 - 04:09 am.

Quoting Lichen on his three day old Honey Tek jar: "as of what--three days, now? it is developing a yellowish sort of stuff in the bottom....looks like sediment. I shake it up and it swirls around like a snowstorm paperweight, then settles again."

My reply is:

"See this is the problem with honey. Honey contains "hidden" impurities: wax and protein. When you cook honey the protein denatures, just like cooking the whites of an egg. The protein is not visible in fresh honey, cook it and the protein denatures and coagulates... Hence sediment. The wax, which is emulsified in fresh honey, separates out upon heating and forms scum... Some of it binds to the coagulated protein, some of it floats to the top. Then there is the color, which varies from honey to honey. It all makes it difficult for people new to liquid culture to determine off colors and contams. I am rapidly becoming motivated to do some experimental research with clear Karo syrup, which is available right on the shelf of any decent grocer... "

And so I did. Here is the proven Tek:

Use 1 Level Teaspoon of Light Corn Syrup (Karo Syrup or the "no-name" store brand of Light Corn Syrup) per 100 ml of purified water. Mix it well to distribute the heavy syrup into the water.

You can use an empty syringe to measure out 100cc, many jars are graduated


The Karo jar is made up and Sterilized with a solid dome lid, inverted so that the seal side is up (we don't want to can it) and the band screwed on loosely. Foil Cover of course.

To shoot the jar I place it in the Glovebox, remove foil, band, and lid, and shoot directly into the liquid. I pre-sterilize some extra jar lids wrapped in foil and I open one in the glove box and place the clean dry lid on the jar mouth with the seal side _down_. Screw the band on tight and replace the foil cap.

To Clone you open the jar and drop in (or shoot with 9er Tek) a sample of macerated tissue. I use the same basic tek when working with spore prints: open the jar, tip the print over give it a flick or a scrape... Plenty of spores drop, rehydrate, and germinate... Incubate, in a 7-10 days you are ready to shoot dozens of jars... Bulk Grain... Store the excess in the fridge, it keeps for a good long time.

Karo jars prepared in this fashion can be shaken vigorously without leaking or contam problems. Everytime I open a jar in the glovebox I always replace the dome lid with a clean sterilized one, seal side down.

It's a super Tek that gets around the problems with honey. Karo is cheap, easy, and to the point. The speed blows the doors off of Agar Culture.

I will give up some liquid culture pointers... You only need 100cc or so (I mean that's enough to shoot 100 jars already). And don't let it get thick, it reduces the refrigerated storage life (because all nutrients are consumed). All you need to grow out and shoot is a mass of mycelium about the size of a quarter in 100cc of liquid.

What you are looking for is wispy, puffy, sometimes ropy white stuff that clumps and hangs on the bottom. You should shake and/or swirl the jar on occasion to try to keep the mass broken up. Before drawing up syringes to shoot jars thrash the mycelium with the needle and break it up (it fragments very easily at this stage). The solution in the syringe need not be dense... But you should see plenty of tiny little "floaties" in there... It's all you need.

Shooting Dextrose/Karo Tek early helps in a few ways: it saves time; it prevents heavy clumping which can clog standard spore gauge needles; the residual sugar, freshly macerated tissue, and change of media gives a "hot" inoculation, showing growth in 24 hours under incubation; and the cultures appear to store better when fridged. The longer you hold them the greater the chance of Contam.

This is a good place to drop another tek tip: Make sure the jars and everything are crystal clean... Not that a bit of filth hurts the tek (because it is pc sterilized) but flecks of dirt in the clear liquid culture _look_ like contams... and you never seem to notice them until after the mycelium has started to take off... You will be swirling in front of a good light and all of the sudden there is a flake of verm or a small grain of rice flour that somehow got in there and then it seems to make all the difference, it's very noticeable... Always difficult to identify.

I was embarrased to find a hair and another fleck of filth floating in one of my Karo jars the other day. Of course they are sterile, I forgot to wipe the jars out before filling... You would be surprised at the dust and crap that gets in the jars at filling time if you are not clean... It does not hurt the tek, but motes of dust and stuff look like contams. In fact motes of cotton dust floating look very much like spores germinating at 24-36 hours. As long as the specks are not growing the culture is fine, but with an eye after a jar or two, you will see fine detail under good light.

You can also add some small chunks of clean broken glass to the jar before sterilizing, shaking the jar with the glass in it fragments the mycelia nicely and speeds up the growth.. But it makes it harder to eyeball.

Quoting Karna:

Another good way to break up the mycelium is simply to draw in a syringe and squirt it back out intot the liquid. This usually disperses the myc very well and helps to break up the larger clumps. As for volume. I use 200-250cc (a pint jar about halfway). I shake every day for aeration and to prevent clumping and matting. When its done in about 12-14 days or so, at rest, the mycelium reaches about 3/4 of the way up from the bottom of the jar. This ensures that all 20-25 syringes have ample mycelium. Since the mycelium is actually cultured in the liquid, any syringe will grow out even if there are no visible mycelium clumps (as opposed to say 9ers cloning tek where if you disperse the tissue fragment in too much water, some syringes may have no mycelium fragments and will not grow anything) but the clear syringes usually don't blast off the same way as the relatively cloudy ones.

Hope you find this info helpful! Happy Shrooming!

Nan

Note: Pure Dextrose is still the best imo, and I do prefer it. The formula for Dextrose Tek is 1 Level Teaspoon Powdered Dextrose (malt works too) per 75 ml of water, or 1-1/4 Teaspoons powdered Dextrose in 100 ml water. Dextrose can be obtained from any homebrew supply or on-line at www.ebrew.com Dextrose is also known as Corn Sugar (suprise!). Karo contains some additional sugars (fructose),some salt, and some flavoring (vanilla) generally, so it is not quite as pure as powdered dextrose... But it is still far cleaner than honey IMO.

BTW on the same page of that link you will also find Gypsum and Lime


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OfflineDrink_Punk_Soda
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Re: Opions Please "Liquid Growth Medium Q/A" [Re: ShaMan1988]
    #1599514 - 06/01/03 10:50 PM (13 years, 6 months ago)

Hey all,

Just one quick question- assuming one doesn't own or have access to a pressure cooker, how long should a jar of karo solution be boiled for in a pot?  I've heard that an hour is too long and 20 minutes is too short.. anyone out there with practical experience boiling this stuff?  Thanks in advance!  :grin: 


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InvisibleMycoCakeEater
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Re: Opions Please "Liquid Growth Medium Q/A" [Re: Drink_Punk_Soda]
    #1703310 - 07/10/03 06:15 PM (13 years, 4 months ago)

25-30 min is what i do. Ive just started Karo.


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Invisiblemicro
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Re: Opions Please "Liquid Growth Medium Q/A" [Re: MycoCakeEater]
    #1704318 - 07/11/03 12:22 AM (13 years, 4 months ago)

Refer to my post above.

If you don't have a pressure gague look up the specs on your pressure cooker. If it's 10 psi go with 20-25 mins.

If you live at a really high altitude, you'll prolly have to go just *a bit* longer.

--
Micro


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(Avatar is Maxxy, a character by Mizzyam, RIP)


Edited by micro (07/11/03 12:26 AM)


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