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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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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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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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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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Re: :LIQUID CULTURE - THE EASY WAY: [Re: scatmanrav]
    #4142713 - 05/06/05 02:33 PM (18 years, 8 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.





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Edited by agar (05/06/05 02:51 PM)


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Re: :LIQUID CULTURE - THE EASY WAY: [Re: GnuBobo]
    #4142770 - 05/06/05 02:48 PM (18 years, 8 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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Re: :LIQUID CULTURE - THE EASY WAY: [Re: backupwards]
    #4143036 - 05/06/05 03:42 PM (18 years, 8 months ago)

Baby Bottle, screw ring & nipple are designed to be sterilized by EVENFLO PRODUCTS COMPANY. They will withstand my autoclave & AA 941 Sterilizer. I tested them in both for 30 minutes each at 20 psi.


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Re: :LIQUID CULTURE - THE EASY WAY: [Re: scatmanrav]
    #4143732 - 05/06/05 06:53 PM (18 years, 8 months ago)

Scat....... :thumbup:

You are not drawing attention away from this. Hell, it's all about LC's.

Do you know that life is just SWEET.

Early Friday evening, weather is warm, a big pitcher of Cadillac Margarita?s, a loaf of cheese -garlic French bread, a tossed garden salad, cold slaw, sweet corn on the cob, garlic - butter - shrimp scampi,  meaty pork ribs on the BBQ & a great looking women at your side (wanting you).

It just cannot get better than that. The Club later, some Al Green, Keith Sweat music to dance to??..Time to Party.


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Re: :LIQUID CULTURE - THE EASY WAY: [Re: GnuBobo]
    #4144557 - 05/06/05 11:29 PM (18 years, 8 months ago)

I use RTV high temp gasket silicone for both.


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Re: :LIQUID CULTURE - THE EASY WAY: [Re: seatrip]
    #4144563 - 05/06/05 11:31 PM (18 years, 8 months ago)

Quote:

seatrip said: hi agar,what gauge needle is used? would a 20g sharp one work




18 gauge is best all around. But, 20 gauge will work, if the myc isn't so thick, it is stringy.


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Re: :LIQUID CULTURE - THE EASY WAY: [Re: VALIS]
    #4146557 - 05/07/05 01:13 PM (18 years, 8 months ago)

LOL...... In another life... I posted here as 6T. agar=SixTango.

Yup, you can use a colonized seed/grain to start an LC.

Just make sure the transfer is done in a CLEAN way.... with MINIMAL open air exposure............otherwise - contam farm.


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Re: :LIQUID CULTURE - THE EASY WAY: [Re: VALIS]
    #4146838 - 05/07/05 02:00 PM (18 years, 8 months ago)

I did miss it. Excuse = very hung over today.

To much FUN last night


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Re: :LIQUID CULTURE - THE EASY WAY: [Re: Blue Helix]
    #4152603 - 05/09/05 07:01 AM (18 years, 8 months ago)

You dont need to inject 2 or 3 ml of spore solution. Any amount (so long as it has a few viable compatable spores in it) will work. More - simply gets the action going faster, IMHO.

I certainly don't advocate WASTING spore solution. At the same time, I prefer to use the content of a spore laden syringe at one sitting, so I do not have partial syringes laying around.

Once - you master mush/cult & PRINT or SPORE SOLUTION MAKING you generaly have more spores in print or solution form - than a single person (or small group) could EVER USE.

I often use a SPORALATOR to make spore solution:



1/2 or 1 gallon wide mouth jar (Sun Tea type) - with spigot removed & replaced with an injection / asperation port - that reseals, installed in the hole in the jar - where the spigot was.

Lid altered with a gas exchange filter & install a clip of some sort to hold a cap suspended with a stainless steel treble hook through the caps stem area - inside the container.

Fill container with 1000 / 1500 / 3000 ml water (depending on size of container) & PC the whole container with water inside it 20 minutes @ 15 psi &  allow to cool to room temp.

When ready for use, harvest a large cap (veil about ready to tear), inside a G/B or F/H - swab cap top with h202, remove veil, peirce cap stem area with sterile treble hook, open container & attach cap to clip, which hangs over sterile water & will drop spores directly into it (repeat once or twice or more w/same strain cap).

The result is 1000 / 1500 / 3000 ml of spore solution that can be asperated through the injection / asperation port on the side of the jar via a sterile syringe.

Test first syringe on agar/petri/plate to insure spore solution contains no contaminates & (if not) presto - you now have a capacity to fill 100 / 150 / 300  syringes out of a single spore laden solution container - in aseptic conditions.  :thumbup:


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Re: :LIQUID CULTURE - THE EASY WAY: [Re: IGnosticAbhorI]
    #4152737 - 05/09/05 08:28 AM (18 years, 8 months ago)

Quote:

IGnosticAbhorI said:
Agar....you're crazy lol Killer "Spore-A-Lator" btw :thumbup:
-Gnostic




Just a FUN hobby......... :wink: :tongue2: :stoned:


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Re: :LIQUID CULTURE - THE EASY WAY: [Re: Silly_Cyben]
    #4156485 - 05/10/05 07:33 AM (18 years, 8 months ago)

Quote:

browilliam said: How long can these bottles be stored in the refrigerator? William




Once you have healthy culture growth, it is BEST to stall them, by placing in the fridge - where they can/will last several months.


1 gallon LC on a magnetic stir plate


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Re: :LIQUID CULTURE - THE EASY WAY: [Re: jarroddupont]
    #4813056 - 10/16/05 09:22 PM (18 years, 3 months ago)

Quote:

jarroddupont said:
Okay so if one fills the nipple with silicon to keep "air and water tight"...does this mean LC's don't need air exchange?




I prefer some gas exchange. But, sealed - you will still get myc - just not as thick.


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