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Amazon Shop for: ½ Pint Jars, Agar, Malt Extract, Pressure Cooker

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Invisibleagar
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:LIQUID CULTURE - THE EASY WAY: * 11
    #4141612 - 05/06/05 11:52 AM (11 years, 9 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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Offlinejtk
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Re: :LIQUID CULTURE - THE EASY WAY: [Re: agar]
    #4141640 - 05/06/05 11:59 AM (11 years, 9 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?


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OfflineSammy
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Re: :LIQUID CULTURE - THE EASY WAY: [Re: agar]
    #4141641 - 05/06/05 11:59 AM (11 years, 9 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


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OfflineHippieChick
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Re: :LIQUID CULTURE - THE EASY WAY: [Re: agar]
    #4141649 - 05/06/05 12:00 PM (11 years, 9 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.
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OfflineSammy
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Re: :LIQUID CULTURE - THE EASY WAY: [Re: Sammy]
    #4141651 - 05/06/05 12:00 PM (11 years, 9 months ago)

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

Sam


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Invisibleagar
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Re: :LIQUID CULTURE - THE EASY WAY: [Re: HippieChick]
    #4141684 - 05/06/05 12:07 PM (11 years, 9 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 12:12 PM (11 years, 9 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?


--------------------
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"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 12:21 PM (11 years, 9 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 12:28 PM (11 years, 9 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 12:33 PM (11 years, 9 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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Re: :LIQUID CULTURE - THE EASY WAY: [Re: Lucid_Euphoria]
    #4141890 - 05/06/05 12:49 PM (11 years, 9 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 01:12 PM (11 years, 9 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 01:54 PM (11 years, 9 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..?


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Offlinescatmanrav
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Re: :LIQUID CULTURE - THE EASY WAY: [Re: Aeon01]
    #4142361 - 05/06/05 03:04 PM (11 years, 9 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..
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Offlineblackout
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Re: :LIQUID CULTURE - THE EASY WAY: [Re: Aeon01]
    #4142370 - 05/06/05 03:06 PM (11 years, 9 months ago)

Ratio is about the same, sugar is not 100% sugar by weight but it is probably best to treat it as such, it is better to have slightly less sugar than slightly more in liquid innoculants. Both will have nutrients for the mycelium to feed on, the honey probably has a bit more but has other things floating in it which make it harder to see what is going on. You can boil up honey and then filter it or decant it off the sediment resulting in a clearer jar so you can clearly see growth (and contams). agar already mentioned the need for straining dry malt extract solutions.

I cannot get karo where I live, most people can get honey.
I reckon maple syrup would work too.


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Re: :LIQUID CULTURE - THE EASY WAY: [Re: scatmanrav]
    #4142378 - 05/06/05 03:08 PM (11 years, 9 months ago)

Quote:

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



It helps but I dont bother myself and have no problems. you should shake up the jar well to aerate it after boiling (which drives off the dissolved oxygen). The catch 22 with a filter disc is that you cannot shake it very hard for fear of soaking the filter which will invite contams.


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Re: :LIQUID CULTURE - THE EASY WAY: [Re: scatmanrav]
    #4142713 - 05/06/05 04:33 PM (11 years, 9 months ago)

Quote:

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




As I stated, the method is not optimal. The reason why is lack of gas exchange. To cure that with the BB/LC tek, you simply cut a piece of tyvek in a circle & use rubber bands to hold it around the BB botle. That way.... once in awhile.. you can twist the cap a 1/2 turn loose, to allow for some gas exchange.

Also, once you have healthy myc in a BB/LC, place it in the fridge - to stall it.

I don't use the method. I just wrote the BB/LC tek, for the novices to get a start on LC's.

I use larger glass containers (quarts - half gallon & gallons), with rubber tubing through the lid, silicone sealed down & polyfil stuffed, to allow for gas exchange (as in the pic's below).

I also have a magnetic stir plate & place a stir bar in each container. Then, use the stir plate to agitate the culture, as well as shred it into TINY pieces. So, I don't have the problem of getting the filters wet. If need be, I can also clamp them off.

What is GREAT about the rubber tubing is, you remove the polyfil in one, insert a sterile syringe, tilt the jar, asperate culture out, withdraw the syringe & reinsert the polyfil. No drips, no mess.

Also, with two (2) tubes, when you asperate culture out of one, it pulls air in the other filtered tube. So, you don't create a vacuum in the jar.

With this tubing method & without a stir plate, you simply clamp off the tubing, then shake the hell out of the jar. Then, set the jar upright & remove the clamps.





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


Edited by agar (05/06/05 04:51 PM)


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InvisibleGnuBobo
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Re: :LIQUID CULTURE - THE EASY WAY: [Re: agar]
    #4142723 - 05/06/05 04:36 PM (11 years, 9 months ago)

Agar...I don't understand how the tubes function on the quart jars of LC shown.  Could you give a quick description of how/why they're used?  Or...so the filter won't get soaked?  :confused:


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Invisibleagar
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Re: :LIQUID CULTURE - THE EASY WAY: [Re: GnuBobo]
    #4142770 - 05/06/05 04:48 PM (11 years, 9 months ago)

Quote:

GnuBobo said:
Agar...I don't understand how the tubes function on the quart jars of LC shown.  Could you give a quick description of how/why they're used?  Or...so the filter won't get soaked?  :confused:




We must have been writing - about the same time. See my post right above - your last post.


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InvisibleGnuBobo
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Re: :LIQUID CULTURE - THE EASY WAY: [Re: agar]
    #4142815 - 05/06/05 04:57 PM (11 years, 9 months ago)

wicked.  Thanks. :wink:


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Amazon Shop for: ½ Pint Jars, Agar, Malt Extract, Pressure Cooker

Mushrooms, Mycology and Psychedelics >> Mushroom Cultivation >> Mushroom Cultivation Archive >> Agar and Liquid Culture

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