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Invisibleagar
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:LIQUID CULTURE - THE EASY WAY: * 18
    #4141612 - 05/06/05 09:52 AM (18 years, 8 months ago)



:LIQUID CULTURE - THE EASY WAY:

(how to turn 2ml/cc of spore solution into 20 ??..ten ml/cc syringes - right - cheap & easy.

Assuming you have an empty syringe & another with spore solution.

Go to Kmart, Target or any large discount Mart & buy a 3 pack of EvenFlow plastic baby bottles.

Also buy a small bottle of Karo corn syrup, a tube of silicone sealant & a few glass marbles.

All the baby bottle parts (bottle - screw cap - nipple) will withstand sterilization in a pressure cooker.

Remove the nipple, apply a small amount (1/8th to ? inch) of silicone sealant to the inside (interior) of the nipple, to serve as a self sealing syringe needle injection point. Allow the sealant to cure overnight

Note: the baby bottle has milliliter (ml) graduation marks up to 240 ml (8 fluid ounces).

Add 200 ml of water to the baby bottle (use marks on it - as a measuring point).

Take your empty syringe and aspirate 6 (six) ml of karo syrup into it.

Then inject that into the open baby bottle, resulting in an approximate 3% solution.

Next, add a glass marble to the baby bottle, to serve as an agitator devise - later.

Insert the nipple into the baby bottle (facing down - inside the baby bottle).

Attach the screw ring lid to hold the nipple in place over the mouth of the baby bottle.

:IMPORTANT:
Attach the screw ring loose, to allow steam to escape & pressure equalization during pressure cooking.

Pressure cook the baby bottle (or several) 20 minutes @ 15 psi.

:IMPORTANT:
Allow the pressure cooker to cool to room temperature, otherwise a vacuum can form inside the bottle, if the screw ring is tightened while the bottle is hot..

Open the cooled pressure cooker & immediately tighten the screw rings on the bottle (or bottles).

You now have a sterilized nutrient solution, in which to inject 2 or 3 ml/cc of spore solution into.

Once you inject a spore solution into it, simply incubate it around 78 to 82F in a clean environment.

You should be able to see tiny strands of mycelium (from germinating spores) in the solution within several days. Be patient, as some spores take longer to germinate - than others.

Once you have visible mycelium in the culture, gently agitate in on a daily basis.

Once the mycelium in the culture is fairly thick, it is ready to be aspirated into sterilized syringes, and used an inoculate.

~~~~~~~

FOR THOSE WITHOUT A PRESSURE COOKER:

NOTE: Not as fool/proof - fail/safe as pressure cooking sterilization.

Cut a piece of tyvek in a circle. Use a rubber band to hold it in place over the screw cap of the baby bottle.

Insert the baby bottle into your microwave (be sure the lid is somewhat LOOSE).

Then, microwave heat it until the solution in it - comes to a boil.

Then - stop, wait a moment & do the same thing again. Repeat, several times.

Remove the baby bottle (gently - as the lid is still loose) & allow to cool to room temperature.

Then, tighten lid.

~~~~~~~~~~~~~~


Or, if you want to do gallons, use jars - with filter lids.


--------------------


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OfflineVALIS
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Re: :LIQUID CULTURE - THE EASY WAY: [Re: agar] * 1
    #4146394 - 05/07/05 12:49 PM (18 years, 8 months ago)

When I was searching through liquid culture (LC) techniques a couple months back, I was particularly impressed with SixTango's Easy Liquid Culture Container Tek.

I like the use of baby bottles because of the ease and convenience of the nipple - c'mon, who doesn't love a good nipple?

Anyhow, here's what I did:




Materials - evenflo 4oz glass baby bottle, filter fiber (i.e. polyfill ), glass marble, tubing, scissors.



Snip nipple just below the tip.



Push the tubing through bottom of nipple.



Cut tubing.



Insert filter material into tubing.



Drop the marble in.



Finished product.


Further information for those who don't already know:

Liquid culture is a method where spores, an agar wedge, or mushroom tissue is injected/inserted/placed into a nutritious solution and incubated, for the purpose of inducing mycelium growth, thereby creating a large amount of inoculum that can be used to inoculate cakes or spawn with. The primary benefits of doing this are as follows:

* Lots and lots ( and lots ) of innoculant can be easily created - 1 10cc multi-spore syringe can be turned into 10 jars of LC, and 100mls of LC can create 10 syringes... pretty cool.

* Using LC to inoculate cakes or spawn can vastly speed up colonization - no lag time waiting for spore germination, in addition to the large amount of inoculation points when injected into substrate.

* The LC can be stored over long periods - throw it in your fridge for months.


Following are the beneficial aspects of using a baby bottle as a container for your LC:

* The nipple serves as a self-healing injection point.

* The nipple can be bent over or squeezed just below the filter tube in order to prevent the LC from saturating the filter when shaking or when drawing up solution.

* Easy and quick to prepare - no need to drill or seal.

* Convenient size, and it's nice to have the ml's/oz's right on the bottle.


Other things, not unique to the baby bottle tek, but useful in any LC container:

* The marble is used as means to better stir/mix the LC, and to break up any mycelium clumps that may occur.

* The filter is for gas exchange.


--------------------
Nature is the Technology of the Divine.


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InvisibleAtheist
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Re: :LIQUID CULTURE - THE EASY WAY: [Re: misterdogman] * 1
    #8179488 - 03/22/08 11:40 AM (15 years, 10 months ago)

8 pints = 1 gallon


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Offlinemisterdogman
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Re: :LIQUID CULTURE - THE EASY WAY: [Re: Atheist] * 1
    #8180568 - 03/22/08 05:35 PM (15 years, 10 months ago)

yeah I was trying to educate yodabuds who aint here neway, he said there was 2 pints in a half Gallon...lol. Well if thats the case I own the Gas station on the corner, want to buy some Gas? LOL.


--------------------
While driving my overpowered car around tight corners like a maniac all my friends yell out. Hey man slow down, we might Wreck. I reply, Shut up, I drive like Dale Earnhardt. They all look around at each other and laugh. Then one asks me. Hey wait, isnt he dead? Well yeah he is I said. Then I drive like Junior I mumble, while I promptly slow down.
"Everyday is opposite day. Everything I say is opposite of what you think I am saying. So if I say something and you think I mean one thing it is the opposite of what I am really trying to say. I actually mean another thing other than what you are thinking I am saying. Get it? Good because that's what I meant".
"You may defeat me, you may even destroy me, But you will never CONQUER me"!


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OfflineScavengerType
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Re: :LIQUID CULTURE - THE EASY WAY: [Re: GnuBobo] * 1
    #8180694 - 03/22/08 06:16 PM (15 years, 10 months ago)

interesting. My LC method isn't spectacular so perhaps this is a good idea for me.


--------------------
"Have you ever seen what happens when a grenade goes off in a school? Do you really know what you’re doing when you order shock and awe? Are you prepared to kneel beside a dying soldier and tell him why he went to Iraq, or why he went to any war?"
"The things that are done in the name of the shareholder are, to me, as terrifying as the things that are done—dare I say it—in the name of God. Montesquieu said, "There have never been so many civil wars as in the Kingdom of God." And I begin to feel that’s true. The shareholder is the excuse for everything."
- Author and former M6/M5 agent John le Carré on Democracy Now.
Conquer's Club


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OfflineOwMyHead
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Re: :LIQUID CULTURE - THE EASY WAY: [Re: Primate] * 1
    #8712987 - 08/01/08 04:06 PM (15 years, 5 months ago)

I'm so glad I read all this thread, because I do, in fact, have bottles with tiny little holes at the base of the nipple (not at the protrusion, but the lip that sits under the cap when you screw it on.

Perfect solution:  a teeny little drip of silicone, and a q-tip.  No muss, no fuss, no mess, and it doesn't keep the bottle from sealing because you have too much gunk on the lip. :smile:


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OfflineAnnoA
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Re: :LIQUID CULTURE - THE EASY WAY: (moved) [Re: agar] * 1
    #9171545 - 11/02/08 08:29 AM (15 years, 2 months ago)

This thread was moved from Mushroom Cultivation.

Reason:
-> archive


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Offlinejtk
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Re: :LIQUID CULTURE - THE EASY WAY: [Re: agar]
    #4141640 - 05/06/05 09:59 AM (18 years, 8 months ago)

Hey agar nice tek. I just did a karo LC and used 5 ounces of water to about 6 ml karo, or a little over a teaspoon. Is that too much karo?


--------------------
Don't let your brain tell your mind it can't feel


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OfflineSammy
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Re: :LIQUID CULTURE - THE EASY WAY: [Re: agar]
    #4141641 - 05/06/05 09:59 AM (18 years, 8 months ago)

Hands down.. a GREAT way to do LC!! I'm going to Targyworld today to pick me up some supplies!! Agar... thanks again for your contri. These pics + great explanation make this project easy even for a nEwB

Samuel


--------------------

I believe in the Golden Rule ? The Man with the Gold . . . Rules.
- Mr. T


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OfflineHippieChick
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Re: :LIQUID CULTURE - THE EASY WAY: [Re: agar]
    #4141649 - 05/06/05 10:00 AM (18 years, 8 months ago)

Nice Tek. The silicon fills the inside of the nipple, right. You don't need to PC these and filter through two coffee filters and then re PC?
Peace,Love,Happiness and Harmony.
:heart: Hippie Chick  :mushroom2:


--------------------
Peace,Love and Happiness
:heart: HC :mushroom2:

Freedoms just another word for nothing left to lose..............

I LUV My Greenhouse
http://www.shroomery.org/forums/showflat.php/Number/5545848#5545848

My First Pans
http://www.shroomery.org/forums/showflat.php/Number/6212058#6212058


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OfflineSammy
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Re: :LIQUID CULTURE - THE EASY WAY: [Re: Sammy]
    #4141651 - 05/06/05 10:00 AM (18 years, 8 months ago)

JTK: I used 1 tablespoon of Karo for my 1/2 pint jars of LC.. it's worked great.

Sam


--------------------

I believe in the Golden Rule ? The Man with the Gold . . . Rules.
- Mr. T


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Invisibleagar
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Re: :LIQUID CULTURE - THE EASY WAY: [Re: HippieChick]
    #4141684 - 05/06/05 10:07 AM (18 years, 8 months ago)

Quote:

HippieChick said:
Nice Tek. The silicon fills the inside of the nipple, right. You don't need to PC these and filter through two coffee filters and then re PC?
Peace,Love,Happiness and Harmony.
:heart: Hippie Chick  :mushroom2:




I could make a LC, just looking at that AVI. (JOKE)

This formula is karo, it doesnt get much gunk in it - when you PC.

Sealing the nipple with silicone simply makes it AIR & WATER TIGHT.

It adds another protective injection port layer, that RESEALS when you pull out the needle.

So, no need to cool, drain, strain & re/PC, as with light dry malt & dry dextrose. Which, I believe is a better nutriant solution. I just did this one with Karo, so the new hands could get an LC togather - QUICK & EASY. It is not optimal, but works well enough to produce plenty of LC myc.


--------------------


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InvisibleLucid_Euphoria
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Re: :LIQUID CULTURE - THE EASY WAY: [Re: Sammy]
    #4141699 - 05/06/05 10:12 AM (18 years, 8 months ago)

Great and simple way to do LC there Agar. I'm gonna give this a try in a month or two on my next grow. I do have a question about LC since I've only read minimally about it. How long can you keep the LC before it becomes dormant/unusable? Also how should it be stored for keeping? I'd assume you don't want to keep it at incubation temps once you've gotten a prolific mycellium culture?


--------------------
PAN CYAN & AZURE FOR TRADE

"If ignorance is bliss, THEN KNOCK THE SMILE OFF MY FACE!" - Zach de la Rocha

"Today a young man on acid realized that all matter is merely energy condensed to a slow vibration, that we are all one consciousness experiencing itself subjectively, there is no such thing as death, life is only a dream, and we are the imagination of ourselves. Here's Tom with the weather!" -Bill Hicks-


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OfflineHippieChick
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Re: :LIQUID CULTURE - THE EASY WAY: [Re: Lucid_Euphoria]
    #4141748 - 05/06/05 10:21 AM (18 years, 8 months ago)

I've heard they stay good for months if stored in the fridge. They don't need to be warmed up before using.
Peace,Love,Happiness and Harmony.
:heart: Hippie Chick  :mushroom2:


--------------------
Peace,Love and Happiness
:heart: HC :mushroom2:

Freedoms just another word for nothing left to lose..............

I LUV My Greenhouse
http://www.shroomery.org/forums/showflat.php/Number/5545848#5545848

My First Pans
http://www.shroomery.org/forums/showflat.php/Number/6212058#6212058


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Invisibleagar
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Re: :LIQUID CULTURE - THE EASY WAY: [Re: HippieChick]
    #4141788 - 05/06/05 10:28 AM (18 years, 8 months ago)

The lovely LADY heard RIGHT.


--------------------


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InvisibleLucid_Euphoria
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Re: :LIQUID CULTURE - THE EASY WAY: [Re: agar]
    #4141810 - 05/06/05 10:33 AM (18 years, 8 months ago)

Exccccelent!
I'm just finishing my first grow now (about to birth my bulk tubs in about 5 days).

Then I gotta put all my stuff away until we renew our lease in a month and a half so if Mr. Landlord decides to do a once over I don't have to explain all the odd ghettofied science lab shit. After that though I'm definately gonna make a couple LC jars and do a second bulk grow. Gonna try out the Equadors next time around.

Thanks for the info.
Catch ya on the flipside!


--------------------
PAN CYAN & AZURE FOR TRADE

"If ignorance is bliss, THEN KNOCK THE SMILE OFF MY FACE!" - Zach de la Rocha

"Today a young man on acid realized that all matter is merely energy condensed to a slow vibration, that we are all one consciousness experiencing itself subjectively, there is no such thing as death, life is only a dream, and we are the imagination of ourselves. Here's Tom with the weather!" -Bill Hicks-


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Offlineblackout
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Re: :LIQUID CULTURE - THE EASY WAY: [Re: Lucid_Euphoria]
    #4141890 - 05/06/05 10:49 AM (18 years, 8 months ago)

Nice guide, I like the fact the baby bottle has the graduations already on it. Some may ask why you put a filter on the microwaved bottle, it is because while cooling it will suck air in through the loose threads on the bottle. If I do not use a filter I always make sure to leave it in the microwave overnight without opening it at all so the air it does draw in will be pretty much free from contaminants. Wipe the microwave down with disinfectant before you use it (you should do this regularly anyway).
Also instead of stopping and starting the microwave you can leave it full until it starts to boil and then turn it down to defrost for 15-20mins. Remember in the microwave you will be boiling off liquid so you need to add more water and less karo/honey at the start since it will get concentrated more as the water boils off (should stay the same water content in the pc).
I would also recommend shaking the bottle well after it has cooled to aerate the mixture with oxygen which has been driven off by boiling. Mycelium will grow better in aerated liquid


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Invisibleagar
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Re: :LIQUID CULTURE - THE EASY WAY: [Re: blackout]
    #4141974 - 05/06/05 11:12 AM (18 years, 8 months ago)

:thumbup: :wink:


--------------------


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OfflineAeon01
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Re: :LIQUID CULTURE - THE EASY WAY: [Re: agar]
    #4142126 - 05/06/05 11:54 AM (18 years, 8 months ago)

blackout you mentioned using honey as a substitute to karo.. this liquid culture procedure is very interesting and i was wondering what the difference between honey and corn syrup is, both in terms of mycelium development and ratio, and the differences between corn syrup and honey..?


--------------------
"You took too much man.. Too much.. Too much.."


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Offlinescatmanrav
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Re: :LIQUID CULTURE - THE EASY WAY: [Re: Aeon01]
    #4142361 - 05/06/05 01:04 PM (18 years, 8 months ago)

How does the baby bottle breathe? Is it unnessesary for gas exchange on LCs?


--------------------
"life is like a drop of rain getting closer and closer to falling into a lake, and then when you hit the lake there is no more rain drop, only the lake."

Growing with bags, start to finish (including my new grain and substrate prep)
Anyone looking to start bulk tubs/mono tubs/shotgun hybrids? Good tubs to use..
How I do grain (old still good tips)
Turn your closet into a fruiting chamber
Casing layer colonization and overlay


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