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Re: Verum's Liquid Culture tests [Re: Tormato] * 1
    #26159272 -

Hey tormata, just wanted to thank you for that tomato lc tek. I had no clue you were such a great mycologist.
:douchewink:


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Masters and slaves hunting clones        How not to isolate (it worked, but)


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Re: Verum's Liquid Culture tests [Re: verum subsequentis] * 1
    #26159287 -

:canthelpbutlaugh:

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Re: Verum's Liquid Culture tests [Re: verum subsequentis]
    #26159307 -

verum subsequentis said:
Hey tormata, just wanted to thank you for that tomato lc tek. I had no clue you were such a great mycologist.
:douchewink:


That shit changed the game for real!!

:bathtub40lol:


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"Lately it occurs to me what a long, strange trip it's been." ~ Grateful Dead

Before you start...Do you have a Pressure Cooker and a Dehydrator? I highly recommend getting both!

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Re: Verum's Liquid Culture tests [Re: Tormato] * 1
    #26162658 -

https://www.shroomery.org/forums/showflat.php/Number/26152975#26152975

updated post below op with some new shit. I had willy myco over for some beers last night and we decided to do some funky coco water experiment. Along with some regular LCs that'll get put on stir plates.


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Masters and slaves hunting clones        How not to isolate (it worked, but)


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Re: Verum's Liquid Culture tests [Re: verum subsequentis] * 1
    #26162762 -

Updated op with
Edit 8/31/19
The time has come to test these suckers on agar.

Typically i just flame sterilize a loop, dip it in the LC and streak a plate. But I've decided to do things as tippy top as possible for this write up for the sake of SCIENCE. I'll also be going rather overboard on the wrapping and labeling. I don't usually do things this detailed and carefully but i want to set a good example for people just learning the ropes and show the safest and "best" way of doing certain things. Like individually wrapping all the needles in foil and what not.

Lately I've been free pouring the LC into bags but here i will fill 60cc syringes for the sake of accurately dosing the bags. I'll take a drip from the syringe, put it on the sterile loop and then streak the plate.

So, now to sterilize everything necessary.
4 60cc syringes
4 16 G. needles
One long ass aspiration needle
Loop
Scalpel (not needed for this but I'll be using it for other things in the session.)



It's good to individually wrap everything you'll need so that you can leave things safely wrapped in foil until you need them.



When wrapping i like to be careful to wrap in such a way that they are protected but will also be easy to unwrap without contaminating. It can also be helpful to label things with a sharpie. You can be as detailed as you want but i just like to know what is what and what end of that thing matters most. This can be done by drawing and arrow, a star, a drawing of the item... Bottom line is your gloves are not sterile. They are only pretty clean. Some items don't matter as much as others. Like the scalpel and loop. They can be taken out all willy nilly and then flame sterilized later. The long ass needle and syringes are a different story though. Those are labeled in such a way to show me what end should be opened first and held with my hand while the other end is carefully and cleanly removed from the foil and dealt with appropriately. Point in case, the syringes show me what end the needle goes in. This is the important end. If you pull the sterile syringe out of the foil and then allow some bacteria or nasty ass spores to make there way in to the end.... You are Fucked.

It's also worth noting that I'll be drawing four different LCs into four different syringes using the same aspiration needle. I'm only doing this because i'm choosing to be all detail and use syringes. I don't want to aspirate with the 16 G needles because that puts my hand closer to the LC. With the long needle I can aspirate without having to have my hand over the LC. Two of these are in half gallon masons and pretty deep. It's easy enough to safely switch out the needle and move on. But I'd highly recommend just testing with a loop and then free pouring.



Then it's good to wrap all your random shite in foil for a few reasons. This pouch holds everything together in the cooker and allows an extra layer of protection while moving the shite out of the cooker and into the flow or SAB. The pouch can be opened and the items removed safely once everything is ready.



Then we cook. This cooker also has four new LCs in it. I also have two jars of nothing with lids on them that i use to safely hold delicate things (like the lid of the LC i'm testing) up in the flow and away from the nasty table. Additional there are a couple jars in there that a simply there to keep my foil pouches from falling in the water.



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Masters and slaves hunting clones        How not to isolate (it worked, but)


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Re: Verum's Liquid Culture tests [Re: verum subsequentis]
    #26164064 -

Updated op with some shit that'll be rather wordy and potentially boring for ya'll experienced mofos but.... I do it for the noobs. These are the types of posts that i loved the ever living shizzle out of when i was learning... So i try to return the favor.

Edit 9/1/19

Last night i filled a filled the syringes and tested these LCs on agar. I tried my best to tape my phone to the ceiling and leave it dangling at just the right angle to get some pics. I filmed the whole session and then went back and took screen shots of some pertinent moments. I tried to crop in and get good examples. You'll probably need to expand the photos and look closely if you want to understand what's going on. Hopefully this is helpful to some people.

I like to move everything to one side of the flow, iso my hands and arms, put on gloves and iso them, get a new paper towel soaking wet and use it to clean off the empty side of the flow table. I'll then clean everything one by one and move it to the clean side and iso the now empty dirty side. I'll then spread everything out and do a mental run through of what I'm about to do. In this picture everything is cleaned with iso and I'm getting my game plan straight.



I can't stress enough, to those of you who are new to sterile sessions, it is very important to know what you want to do and how you want to do it. I've done thousands of sterile sessions and i still run through my procedure in my mind before i begin. I then organize all items in the best way I can think of to facilitate smooth maneuvering throughout the session. This pic is post intentional and strategic organization of the items.



Now I'll start taking my shizzle out of the foil pouches. Always being mindful of what is most important, what is least important and what doesn't fucking matter at all.

Syringes



Scalpel needles and loop
All individually wrapped as to remain nice and sterile until it is their time



Here's where the rubber meets the road. Unwrapping things that must remain sterile until done, one must be mindful of every move.

The stainless aspiration needle will be removed in such a way that the luer lock end remains in the safest position possible. It should always remain in "first air" (i believe is the term). This means nothing should ever touch it or be between it and the filter. This is one reason labeling your foil can help. I know which end is which without having to fuck around.

   

Then i go about unwrapping a syringe. Keeping in mind that the luer lock end of the syringe is of upmost importance and must remain in "first wind". All the while keeping the luer end of the needle in a safe position.

 

Now, as i connect the needle to the syringe i keep both luer ends in first air.



Flame the needle. I was careful not to touch the needle too close to the luer end because i didn't want to have to flame it all the way down. Doing this can melt the end of the syringe an cause all manner of problems.

 

Then i swirl the LC, remove the lid and set it upside down on the lid of one of the sterile jars that just came out of the cooker. Then tilt the LC and aspirate.



Then i set the syringe on the rack in such a way that every part that matters is floating in first air. The open LC is also very vulnerable and must be respected. Now we retrieve the lid and carefully place it back on the LC.

 

Now we streak. It isn't entirely necessary to re flame the needle but is definitely a good idea. You'll have to flame both the needle and the loop and then allow both to cool without compromising either. It's a little odd, learning how to hold things at first but gets easier with a little practice.




Once cooled, place a single drip on the loop. If you hear any sizzling it wasn't cool enough.



Safely set your syringe down without compromising your precious innoculant that is suspended in the loop.



Remove the lid of the petri without contaminating it. Finger placement is important. Then streak the LC across the plate. I just zig and zag from top to bottom. Probably 2 zigs and 3 zags.

   

Then i wrap the petri and LC lid with glad cling wrap. Some use parafilm but i like glad.

   

Now I remove the stainless needle and replace with sterile 16 gauge needle. Always remember what matters and what doesn't. Right now the business end of the needle and syringe are of utmost importance.

 

Always label as you go or you'll get corn fused later on.... Fo Sho

 

I like to put syringes into brand new ziplock bags. Just to be safe and sound until ready to use.



Generally, within two days i can tell if the plate is clean but the longer i wait the more obvious it is.


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Masters and slaves hunting clones        How not to isolate (it worked, but)


Edited by verum subsequentis (09/02/19 03:04 PM)

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Re: Verum's Liquid Culture tests [Re: verum subsequentis]
    #26164066 -

and i immediatly see typos. I fucking hate typos. Proof reading time


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Masters and slaves hunting clones        How not to isolate (it worked, but)


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Re: Verum's Liquid Culture tests [Re: verum subsequentis]
    #26164445 -

kick ass. excellent work, bruv. thank you.

in your considered opinion, would you say lc is a strictly large scale process, or does it have value for small scale projects as well?

been doing rice lately, which in order to make it easily shakeable has to be fairly under-hydrated. been thinking maybe using lc to inoculate would be a convenient way to introduce extra water following sterilization, if it turns out the under-hydration causes other less obvious problems later.


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the j stands for jesus.
2020 new years grow along

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Re: Verum's Liquid Culture tests [Re: J. Jack Flash]
    #26164550 -

LC could be used for small grows but going big is where it comes in really handy. I'd say just going agar to grain is the safest, easiest, and most reliable way for growing a few tubs here and there. I've never used rice but I think that's why we don't use it. I use WBS. It's easy, cheap and reliable.


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Masters and slaves hunting clones        How not to isolate (it worked, but)


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Re: Verum's Liquid Culture tests [Re: verum subsequentis] * 1
    #26164797 -

This sort of thing is why I love this forum. Top quality.

Appreciate the effort my man!

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Re: Verum's Liquid Culture tests [Re: Zakkery]
    #26164806 -

Same. Loving all the photos and pointers. Felt like I was in the room.watching you do it as I read along. Very helpful. Very educational. Thanks bud.

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Re: Verum's Liquid Culture tests [Re: Bph]
    #26165152 -

Excellent posts can't wait to see the growth on the plates! Never done LC, but living vicariously through your post 

:notreadyforthis:


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Re: Verum's Liquid Culture tests [Re: Dispsiple]
    #26165216 -

:hi:


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If I ever give out misinformation please inform me so I can have the correct information. :cheers:

Tmethyl said:
Chuck Norris once roundhouse kicked a monotub that wasn't pinning fast enough. The force of the kick rearranged the genetics of the mushrooms, we now call them Penis Envy.

Caps McGee said:
:thumbsup:
Fun part is figuring out what works best for you

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Re: Verum's Liquid Culture tests [Re: verum subsequentis]
    #26165268 -

verum subsequentis said:
I'd say just going agar to grain is the safest, easiest, and most reliable way for growing a few tubs here and there.


Idk... I'm getting 50-60 healthy spawn bags a week, currently, from A2G and G2G. I think its a great option for growing a shit load of tubs, myself. :shrug:

LC is also a powerful tool.


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:mushroom2: LotKid's PE Tips        :shpongle:    LotKid's Chocolates :mushroom2:
:dancingbear: :dancingbear:

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Re: Verum's Liquid Culture tests [Re: J. Jack Flash]
    #26165270 -

J. Jack Flash said:
been doing rice lately, which in order to make it easily shakeable has to be fairly under-hydrated. been thinking maybe using lc to inoculate would be a convenient way to introduce extra water following sterilization, if it turns out the under-hydration causes other less obvious problems later.


I found adding popcorn to rice makes it shakeable, its like ball bearings in a paint can. Popcorn should be heat treated for longer so I have some I did stored in the freezer which can be added to rice jars. I guess marbles would do the same thing but be careful not to shake too hard.

Not only does adding LC introduce water it can add enough myc that no shaking will be necessary as you can get total coverage of myc. I have had jars colonise in just a few days by adding shitloads of LC, calculated amounts. I have found that it is best to hydrate grains fully, or even overhydrate and then dry them out before going in the PC. This way they have been expanded and are like sponges ready so soak up the LC. If you are doing brown rice and purposely not hydrating much in the first place it will not take up as much liquid, but still might do OK. Grains can be dried in the oven or microwave. I have dried grains out so they were drier than they were to begin with, these then had LC added little by little -that was just an experiment and can be dangerous in a microwave as it can cause a fire. If drying in a microwave always have another jar of water so if the grains do go dry the microwaves can at least go into the water.

By weighing jars you can monitor moisture loss. To get nice thick myc I use big syringes, suck in say 60ml and leave the syringe to settle, needle side up for a few hours or a day -this leads to the myc collecting in the bottom. With the needle still upwards I squirt it out over a basin or in the shower, so all I am injecting in jars is the thicker myc at the bottom.

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Re: Verum's Liquid Culture tests [Re: J. Jack Flash]
    #26165303 -

J. Jack Flash said:
in your considered opinion, would you say lc is a strictly large scale process, or does it have value for small scale projects as well?




Lotkid, stay outta here with your G2G bullshit.
:clintonshuh:

I totally understand that G2G can produce a shitload of spawn and that you roll like a champ. BUT, that's not what he was asking.

And Blackout, your post just boggled my brain in multiple ways


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Masters and slaves hunting clones        How not to isolate (it worked, but)


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Re: Verum's Liquid Culture tests [Re: verum subsequentis]
    #26165310 -

:takingnotes:


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Wide Mouth 1/2 Pint No-Pour TEK
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Re: Verum's Liquid Culture tests [Re: verum subsequentis]
    #26165349 -

verum subsequentis said:
J. Jack Flash said:
in your considered opinion, would you say lc is a strictly large scale process, or does it have value for small scale projects as well?




Lotkid, stay outta here with your G2G bullshit.
:clintonshuh:

I totally understand that G2G can produce a shitload of spawn and that you roll like a champ. BUT, that's not what he was asking.

And Blackout, your post just boggled my brain in multiple ways


I was just going off your sentence. In that I feel A2G and G2G are very viable and reliable options to grow more than just a few tubs here and there. That's all. :grin:


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:mushroom2: LotKid's PE Tips        :shpongle:    LotKid's Chocolates :mushroom2:
:dancingbear: :dancingbear:

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Re: Verum's Liquid Culture tests [Re: LotKid]
    #26165368 -

I know, and you are right (as you know). I'm running some tests now to try and see if G2G can compare with LC in all aspects. Hours/ effort/ time/ speed.... you know. Normally I make up a huge LC, noc it up in a quick flow session (literally takes just a minute or two), test it quickly and easily by dipping the flamed loop in and then use it on around 40 gallon bags.

My last test showed that a wbs bag with only 1/10th of a myco quart added colonized nearly as quickly as one with half a myco quart and both colonized faster than the bag with around 15cc LC free poured in. But i didn't shake the LC bags when i should of and think they could've been roughly on par if i had.

I'll do some G2G when i use these LCs and document the rate of growth but, as anyone who's grown much magic knows, it takes more than one test to really prove something with this many variables


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Masters and slaves hunting clones        How not to isolate (it worked, but)


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Re: Verum's Liquid Culture tests [Re: Just_A_Noob]
    #26165380 -

blackout, that's counterintuitive. to me anyway. i'd think underhydrated grain would more readily absorb moisture than fully hydrated grain. so you're suggesting fully cooking, then fully drying before sterilizing, then inoculating and rehydrating with lc? it's madness.

also, bully on the popcorn for agitation idea. i thought of using marbles, but like you said, seems unwise to vigorously shake a cheap glass jar full of marbles. i can see popcorn working well for this.

re: lots and lots of tubs... maybe in retirement. many years to sharpen my skills and processes before then. the brown rice thing for me started only because i already had it and oats, and the pan cyans i have had a hard time on oats. tried rice just to see, and it colonized super stupid fast. then, the fruiting performance wasn't so great, but i never ran cultures from that project on oats, so had nothing to compare it to. jury is still out. currently running a side by side comparison of some pan cinctulus between wbr and oats.

back to lc, verum again in your opinion, would you consider doing lc in a sab a reasonable practice, or is a hood pretty much required?


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the j stands for jesus.
2020 new years grow along

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