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Agar Jars-->Add Grain-->G2G/GLC-->Expand -The Power Of The Small -


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#1 hyphaenation

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Posted 12 February 2021 - 02:38 PM

Using The Phorce With Small Agar Jars

In recent memory I've had a few disappointing runs where I wanted to dust off my prints and grow some fruits. I started on the wrong foot , even though I knew better. Not sure if laziness or  not thinking it through was culprit. Anyhow I started many jars of grain and inocced a good amount of spores....nothing. Week later started another batch but let the spores hydrate longer , shrunk the size of the grain jars a bit and tried again. Long story short I ended up wasting time, money, supplies and a whole bunch of my patience. Luckily I got no trich! But I also got no redboy myc either for all my efforts.

I decided to go back to the drawing board and stuff I learned here.

I knew already that starting spores on agar is a better bet especially for someone like me who is far less then sterile at the best of times. Some of this is stuff I have shared in the past and its just kind of streamlined a bit. I try and use things I can find in my house wherever possible and try and keep it on the simple side.

I started with 150 ml small glass jars and made a small breather for each. I used a simple potato dextrose agar recipe and stuck with it. Weighed all ingredients and followed procedures. Loaded the small jars and removed from PC and put in clean work space.

From there I tried a few technique of getting the spores to germinate. First i squirted the smallest drop from a spore syringe that I could into the agar jar , let it sit an hour and the swirled and set upside down. The other way was actually flicking flakes of spores off of the tinfoil with a clean implement. They just kind of scrape off onto the agar.

The spores I was using are dated from more than 10 years ago. that's likely why they had such a hard time waking up on the grain jars. Even with the agar many did nothing. Actually 11 out of 12  of the original attempt did nothing and I was ready to give up until I looked in that last agar jar. There it was white mycelium!

As that myelium grew out a bit I added sterilized grain (popcorn) and the myc immediately jumped on to the grain. When that colonized I was ready to start expanding...
 

post-104353-0-07214400-1613157692_thumb. Start spores on agar in small jars (or whatever you prefer)

 

post-104353-0-89401100-1613157699_thumb. When spores germinate and colonize add sterilized grain

 

post-104353-0-36948900-1613157708_thumb. Taller jar is even better for more headroom to add water/shake etc

 

post-104353-0-11083500-1613157717_thumb. A simple way is to G2G the colonized grain into another grain sterilized jar

 

post-104353-0-40827700-1613157732_thumb. The growth from G2G is very quick
 

post-104353-0-87143100-1613157747_thumb. All done in a few days. Ready to be milked

Building Mycelium Milker

I made up this tool with only on-hand gear from in the kitchen... Here were the basic steps I took to build my GLC milker. This tool is used to strip the mycelium from the colonized grain and then that liquid mycelium is used fresh to inoculate.

Guerilla Glue  2  small jar lids together top-to-top (allow to dry properly)
Clean up/scrape off exta glue that fluffed out
Add hose-clamp

Drill holes in jar lid to allow mycelium to freely flow with the water
 

post-104353-0-00585100-1613158407_thumb. Basic mycelium milker parts
 

post-104353-0-86006400-1613163169_thumb. Guerilla Glued the two lids together and let dry, cleaned extra glue off

 

post-104353-0-33139600-1613163178_thumb. Milker ready to go with hose-clam , filter-wheel and assembly

 

post-104353-0-93248600-1613158417_thumb. Gluing done and hose-clamp added

 

post-104353-0-50622000-1613158427_thumb. Rubber band goes around the milking filter-wheel. It keeps it in the milker instead of falling out

I needed to come up with a way to keep that filter-wheel inside the milker and not have it stay behind on the jar when you lift the milker. This little mod works perfect.
 

post-104353-0-59319800-1613158440_thumb. Milking filter-wheel in place. It won't remain on the jar when milker lifted off.

 

post-104353-0-84299500-1613158449_thumb. Grain side of the milking wheel. Rubber gasket for better seal

The milking wheel is sterilized inside a PP5 tupperware with the lid slightly cracked  to avoid vacuum-effect.

post-104353-0-57311600-1613158622_thumb.


It gets placed on to the colonized grain jar (for this photo it is just normal popcorn, not colonized)


post-104353-0-15739800-1613158470_thumb.

That gets flipped over and then secured on to the jar with the water. I sterilize my water in PC and let cool. Pop the lid and then attach the grain jar/milker. Tighten that up snug on both connections, tweak the hose-clamp a turn to keep that tight.
 


post-104353-0-48620200-1613158494_thumb.


Flip that over and now the water rains down into the colonized  grain jar which you bashed to break up the grain mass. Swirl, shake , agitate this and turn upside down. All mycelium water rushes out of the grain. I repeat and then let it sit to drain properly, even shaking it lightly  to get last drops out.
 

post-104353-0-33612700-1613158460_thumb.

 

Now (like 2 pics above) the drained grain jar is above attached to the milker and both can be removed. A lid is placed on the fresh GLC and grain jar is flipped, milker remover and lid with breather placed on it. The GLC can be used in whatever way you like now. I find it a bit thick for any syringes I have so i opted to make a GLC Dabber"
 

post-104353-0-83734600-1613113352_thumb. GLC dabber poots out 3 CC every time you "flick" it

These larger jars of popcorn can take up to 10 cc of liquid before maxing out ... so I tried an experiment. I dabbed half the jars with 6 cc (2shots) and the other half with 3 dabs (9 cc). In my clean area I simply lifted the lid on each jar a crack and dabbed in a poot of the fresh GLC milk.

After 24 you don't see any growth and its disappointing and you think to yourself "well I can always add some colonized grain for a G2G..." but then you check in the morning 36 hours after inoc and there it is ...everywhere. The jars are crawling with inoc points in all sectors. Have not noticed big differences in the 6 cc jars compared to the 9 cc inoc'ed jars. Will update.

 

 

post-104353-0-64736200-1613158765_thumb. 36 hours after inoc with the dabber

 

post-104353-0-48448100-1613158775_thumb.


I will continue to edit and update as I have time.

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Edited by hyphaenation, 12 February 2021 - 04:06 PM.

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#2 Microbe

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Posted 12 February 2021 - 03:00 PM

Im speculating that the lid contraption you made, works similar to an hour glass? Once the water drains into the popcorn, you swirl it around then invert it again, amd the myc laden water returns to the jar? What is that blue rubber band looking thing and how did you get it to flatten out making what appears to be a gasket? Do you have a link to instructions for making one of those? Im pro GLC and i seen your 'GLC Dabber' and im wanting to so something similar. Im so sick of using syringes. I was looking into micro autoclavable infusion pumps.....that how much im tired of using syringes with my glc.
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#3 EYMAIOS

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Posted 12 February 2021 - 03:40 PM

.

Great Topia Saint-Hyph...

.

We bow!



#4 hyphaenation

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Posted 12 February 2021 - 03:42 PM

Hey buddy! Great to see you. Hope your well.

Yeah you got the idea.. water comes in , swirl myc off of the grain and flip over to drain. Add dabber lid to GLC jar. I just drilled a hole and then inserted a needle tip plastc with the end nipped off. It pressure-fit but i top-glued anyways. When it's on the GLC jar and it is going to be sitting around, say to recover etc then I add polyfil as a breather...else I just have a flat lid on there.

The blue rubber band keeps the filter-wheel inside the milker when you lift it off. If there was no rubber band then that lid would just sit there and you already have both hands occupied. It especially comes into play, and I made it for the purpose of milking several jars in a row and I wanted that filter-wheel to stay in the milker in between jars.

 


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#5 hyphaenation

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Posted 12 February 2021 - 03:43 PM

Maiden voyage for this unit...

 

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#6 Microbe

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Posted 12 February 2021 - 03:57 PM

Hey buddy! Great to see you. Hope your well.

Yeah you got the idea.. water comes in , swirl myc off of the grain and flip over to drain. Add dabber lid to GLC jar. I just drilled a hole and then inserted a needle tip plastc with the end nipped off. It pressure-fit but i top-glued anyways. When it's on the GLC jar and it is going to be sitting around, say to recover etc then I add polyfil as a breather...else I just have a flat lid on there.

The blue rubber band keeps the filter-wheel inside the milker when you lift it off. If there was no rubber band then that lid would just sit there and you already have both hands occupied. It especially comes into play, and I made it for the purpose of milking several jars in a row and I wanted that filter-wheel to stay in the milker in between jars.

All is good and like wise. Im going to make one of those. I have been using plates to inoculate jars then g2g from there but im so sick of pouring plates and making endless transfers to maintain enough inoculate. Im so damn busy these days i need to become more efficient.
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#7 hyphaenation

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Posted 12 February 2021 - 04:00 PM

Working on this other concept at the same time...  GLC on tap! It squirts 3 cc of solution reliably every time!

 

Dabber is PC'able though... and thus-far I can only clean the pump by running cleaning solution through it. I wrecked one PC'ing at 15 PSI. Not even sure I needed 15 PSI but it buggered something in it. If you pumped a bit of soapy/bleachy water through the pump and then a bit of clean water I think it would be fine...but I have to experiment a bit. There might also be ways of protecting it better inside the PC and/or sterilize at a lower temp.

For now I am very happy 'dabbing!

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Edited by hyphaenation, 12 February 2021 - 04:04 PM.

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#8 Microbe

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Posted 12 February 2021 - 04:06 PM

Working on this other concept at the same time... GLC on tap! It squirts 3 cc of solution reliably every time!

Now that is what im talking about. I tried to make something similar for agar, lc, myc water and etc using what was supposed to be autoclavable squeeze bottles such as the kind that tattoo artists used but they melted. I wonder if i could use a empty condiment bottle assuming its PP5, and rig a luer lock attachment to it so i can attach a sharp to it. Man that pump is ideal though. Have you tried it yet?

#9 hyphaenation

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Posted 12 February 2021 - 04:14 PM

Have not tested since I destroyed in PC. Will re-calibrate with a new one and get back to you. Like I said with proper cleaning procedure it should not even need a PC'ing but we shall see. If you get time maybe try it out and let me know.

I know the dapper is dandy for sure and it also puts out 3 cc per poot.

Again.. that dabber tip is just a luer-lock needle tip with its closed end cut off. pressure fit into the hole and glued on top to stabilize.

post-104353-0-56668200-1613164441.jpg

Just 500 mils of water added to make the GLC like above is good for 166.6666 of the 3 cc 'poots with the dabber !

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Edited by hyphaenation, 12 February 2021 - 05:26 PM.

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#10 sandman

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Posted 12 February 2021 - 04:49 PM

Those are really clever hyph.


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#11 hyphaenation

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Posted 12 February 2021 - 04:56 PM

Here is 2 jars of popcorn that were hit with the dabber 1.5 days ago. Whereas there was little noticeable growth after 24 hours ...just several hours later the growth is really coming on.

On the left 9cc triple dab and on the right 6 cc double dab.

 

 

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#12 hyphaenation

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Posted 12 February 2021 - 05:44 PM

Less then 48 hours after getting milked for GLC the popcorn has completely recovered and it's even fluffier then before. It's ready to go again but the first 500 mils was already enough to inoc more then a hundred jars!

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Edited by hyphaenation, 12 February 2021 - 06:11 PM.

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#13 hyphaenation

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Posted 12 February 2021 - 06:06 PM

Remember...

post-104353-0-37594200-1613171186.png

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Edited by hyphaenation, 12 February 2021 - 06:07 PM.

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#14 Microbe

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Posted 12 February 2021 - 07:44 PM

Less then 48 hours after getting milked for GLC the popcorn has completely recovered and it's even fluffier then before. It's ready to go again but the first 500 mils was already enough to inoc more then a hundred jars!

Thats why i fell in love with GLC! Brother that is some fast recovery and ready to go again. Have you ever filled the jar up more then just a small layer to get more mycelium mass?

Edited by Microbe, 12 February 2021 - 07:55 PM.

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#15 rockyfungus

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Posted 12 February 2021 - 07:44 PM

I'm trying to rig something up with old RX bottles, they are pp5 and that lid fits 2 bottles technically.


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#16 hyphaenation

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Posted 12 February 2021 - 08:12 PM

 

I'm trying to rig something up with old RX bottles, they are pp5 and that lid fits 2 bottles technically.

Feel free to share here if you'd like... let us know if you need any help/advice.
 

 

Thats why i fell in love with GLC! Brother that is some fast recovery and ready to go again. Have you ever filled the jar up more then just a small layer to get more mycelium mass?

 

post-104353-0-42966100-1613178322_thumb.


Praise GLC! I am a true believer ...


Yeah I usually will have at least half a large jar of colonized grain to start with...you could do it with any amount really. For cubes this is handy and solves a bunch of my problems with starting properly but one or two 500 mil milk runs and I have way to much. So it's not something needing to be done very often for cubensis. I think what I'll do is put the freshly recolonized , original small agar jar in this thread , now full of colonized popcorn...in a clean box  in a bag and place it in a quiet dark place in my cold basement for storage until I need it. I find they can last many months without deterioration like this.

As far as edibles like oysters , woodlovers  and other species that you want a bunch of mycelium to inoculate spawn but you you don't want to use grain due to possible indoor contam...this is perfect! You can endlessly harvest huge volumes of mycelium that is free of grain.


 

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Edited by hyphaenation, 12 February 2021 - 08:12 PM.

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#17 rockyfungus

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Posted 12 February 2021 - 08:25 PM

Been stashing RX bottles for a minute now. I knew they had a purpose just didn't know what it was. (fuck single use plastics)

 

Here ya go a simpler version of that elegant design.

 

RXlid.JPG

RX.JPG

Just need to figure out if the Vet/pharmacist will give away those huge bottles. These don't leave much room to play with


Edited by rockyfungus, 12 February 2021 - 08:26 PM.

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#18 hyphaenation

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Posted 12 February 2021 - 08:41 PM

That looks cool!

Give a test run with just water when you can (when all glue is completely dry etc)...or a dry run with some grain and water . To practice and observe if you have any leaks etc....

 


Edited by hyphaenation, 12 February 2021 - 08:51 PM.


#19 rockyfungus

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Posted 12 February 2021 - 08:54 PM

Good call on testing it! Really tight seal, drains super slow, tapping speeds the process up. I'll play with the gauge. (Edit: Just tilt bottle and half of the holes are exposed to air, very quick drain!)

I have a bunch of colonized baby jars. I could PC that contraption, pour some colonized grain in the one 1/2, and use the sterilized water. Instead of starting grain in there and waiting it out.


Edited by rockyfungus, 12 February 2021 - 09:12 PM.


#20 Arathu

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Posted 12 February 2021 - 08:59 PM

https://en.wikipedia...eristaltic_pump

 

Might be of interest....if you haven't already thought of it.....

 

A


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