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InvisibleViolet
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Invitro casing layers * 8
    #19552215 - 02/11/14 12:37 PM (10 years, 1 month ago)

The last few months I've been experimenting with using soil casing layers for invitro container grows.
It's gone very well.  Far better than my uncased invitros. And I'm pretty sure I know why.


Almost all of my first-flush fruits on Uncased grains come from the container's plastic or the barrier at the sides of the cake, the places as far away from the dense nutrition of the grains as the mycelium can get, regardless of how ideal the conditions are. Every flush after that still forms from the sides/edges.

A casing layer on those straight grains is like the key that unlocks full fruiting performance for every flush instead of the shy and conservative flushing when uncased.
It's not just the ideal humidity microclimate (because my invitro grains keep that anyway), it's the addition of a no- or low-nutrient surface to pin on.


I've come to realize that Cubensis likes to fruit away from nutrient density when possible, and when it is indeed possible it seems to fruit more and better.
Casing layers accomplish this as well as majorly helping to direct fruiting to the place and direction we intend for it.
Bulk substrates also accomplish this by surrounding nutrient sources with low-nutrient watermass little differently than casing layers do, and can maybe be thought of as a "360° casing" for the grain 'spawn'


Even with the moisture-retaining conditions of invitro, I see FAR FAR FAR better results with the addition of a casing layer!

I believe this explains how my cased grain results are in the same range of performance as "spawning to bulk", since the casing layer provides this condition and bottom-watering provides the required moisture over several flushes.


The pictures below are of a throw-away culture that I definitely won't be using anymore.  But casing the cake for invitro fruiting nonetheless allowed the culture to have a full yield for the first time I've ever seen it.




Compare to this below, literally the Best result of the likes that I've seen uncased (most are worse)



Bottom-watered second flushes coming in.



These grows are essentially NO-maintenance and are giving me close to the results of a wonderfully pulled off "V-tek" grow like I normally do.

I love this method. I am kindof thinking now that this is my grow method of choice.
Brown rice, soil casing layer, room for fruiting in the container, no-maintenance except watering which is SO easy and quick. Even if one doesn't purposefully maintain sterility within the container when casing, the somewhat still-air conditions of invitro make spore printing these caps effectively a sterile procedure.  And of course cloning is close to that as well.

May I reiterate:  NO-maintenance! Except watering twice per flush (with good culture) which doesn't even take as much time as one would want to LOOK at them.
The conditions keep themselves perfectly with a loose lid.  It almost does it perfectly invitro for starters, but the casing layer ensures it.
I haven't misted a single one at all.


The only drawback of this method, compared to the "V-tek" as-per with the pint containers in fruiting chambers, is that these containers take up double the space in the cooker.  My small AA910 can fit 6 pint containers to sterilize for 60-70min, but of course only 3 of these containers to sterilize for 60min.

But the advantage this gives is the complete lack of any kind of fruiting chamber, not to mention the increased reliability and the sterile potential.
As long as one can do a nice clean transfer to inoculate, it's pretty much a Sure-Thing Grow.



Even if one only manages 200% BE from the rice (I often get 300-340% with my good cultures in well-done grows), the ~70g of rice in each container would have yielded ~14g after numerous flushes.  A 2lb bag of rice would yield ~90g in just 6-7 containers.
And if you get 300% BE, 21g per container. 2lb of rice grows 136g.
The culturing-mastery "holy grail" 400% BE, 28g per container.  2lb rice grows 181g.

For me 1lb of rice, prepared and divided into Six quart containers, takes just two one-hour cooker runs with my small AA910.



Soon I'll be doing this with some of my much better cultures.


--------------------
Intentionally or not, here in mushcult we are purveyors of love culture and enlightenment movement. Let's try to act like it!

PODS TEK - Growing Invitro with BRF/verm or Grass Seed containers
The simplest, quickest, safest tek!  For beginners, culturers, lazy people, stealth lovers, contam haters, and alternative seekers!
Violet's Teks and Posts

Edited by Violet (03/01/21 09:15 PM)

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InvisiblebodhisattaMDiscordReddit
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Re: Invitro casing layers [Re: Violet]
    #19552246 - 02/11/14 12:47 PM (10 years, 1 month ago)

:thumbup:

I like it, I actually just bought the 32oz ziplocks to try some in-vitro stuff. I think it will be pretty suitable for the PanCyans.

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OfflineBigGreenMat
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Re: Invitro casing layers [Re: bodhisatta]
    #19552299 - 02/11/14 01:03 PM (10 years, 1 month ago)

Agreed violet. My in vitro Ape wouldn't even pin until I cased them with  with sterilized jiffy mix. It took a long time but they are pinning and fruiting decently now. However they are still side pinning. My rice petri bottom fruited pretty impressively.


--------------------
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OfflineBigGreenMat
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Re: Invitro casing layers [Re: BigGreenMat]
    #19554615 - 02/11/14 09:21 PM (10 years, 1 month ago)

Violet quick question for you, but what does your jiffy mix smell like when you put it on the grains?  Do you think it is sterilized by your microwaving?


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My Growth So Far!

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InvisibleViolet
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Re: Invitro casing layers [Re: BigGreenMat]
    #19555031 - 02/11/14 11:27 PM (10 years, 1 month ago)

I use the fine stuff from sifting potting soil.  It smells pretty much like dirt, little different than moist potting soil for a potted plant.
It's definitely not sterilized.  It's not "pasteurized" per se, but that doesn't mean it's not treated as well, just not by the same method.  The goal is the same with dual-microwaving treatment and for me that goal is met every time.  Present contaminants are killed and the defensive bacteria that are missing in unprotected contam-crazy sterilized materials seem to be defending the microwaved casing layer.
The effective and important difference is that this material, combined with this preparation, seems perfect to me in any aspect I can think of.  Fluffy and clumpy, very moist and retaining of that moisture, not at all contam-prone, loaded with microclimate pockets as long as not packed down which it shouldn't be, and not to mention the treatments are much shorter and don't involve tons of highly heated water for a "bath"




--------------------
Intentionally or not, here in mushcult we are purveyors of love culture and enlightenment movement. Let's try to act like it!

PODS TEK - Growing Invitro with BRF/verm or Grass Seed containers
The simplest, quickest, safest tek!  For beginners, culturers, lazy people, stealth lovers, contam haters, and alternative seekers!
Violet's Teks and Posts

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InvisibleViolet
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Re: Invitro casing layers [Re: Violet]
    #19565996 - 02/14/14 11:47 AM (10 years, 1 month ago)

Yet another.  Same throw-away culture.


I can't wait to see this from my good cultures.



Wanna see other people try it too!
It also works with glass jars of course!


--------------------
Intentionally or not, here in mushcult we are purveyors of love culture and enlightenment movement. Let's try to act like it!

PODS TEK - Growing Invitro with BRF/verm or Grass Seed containers
The simplest, quickest, safest tek!  For beginners, culturers, lazy people, stealth lovers, contam haters, and alternative seekers!
Violet's Teks and Posts

Edited by Violet (02/14/14 11:53 AM)

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Offlineblojo02184
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Re: Invitro casing layers [Re: Violet]
    #19566127 - 02/14/14 12:17 PM (10 years, 1 month ago)

So no liner, do you have a problem with side pinning?
And this was rgs or rye berries?

The 2 liter bottle FC or whatever looks like a neat idea.
Would minirounds be suitable?

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InvisibleViolet
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Re: Invitro casing layers [Re: blojo02184]
    #19566227 - 02/14/14 12:49 PM (10 years, 1 month ago)

Of course no liner.  I don't have "problems" with side pins even when they do occur, which is much less when using casing layers.

I don't use anything but grass seed and brown rice for Cubensis and other Psilocybes.  These were brown rice.


Would minirounds be suitable for what?  In my opinion they're only good for agar, and not even that great for that either, so I don't buy them.  For me they're a waste of money.  I vastly prefer ziploc pints for their countless uses and idealness for each of them.
They definitely would not be much suitable for ANY form of invitro fruiting because they're so damn small.


Yeah I like the bottle "chamber" a lot, I have a couple of them over some cased containers at the moment.


--------------------
Intentionally or not, here in mushcult we are purveyors of love culture and enlightenment movement. Let's try to act like it!

PODS TEK - Growing Invitro with BRF/verm or Grass Seed containers
The simplest, quickest, safest tek!  For beginners, culturers, lazy people, stealth lovers, contam haters, and alternative seekers!
Violet's Teks and Posts

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Offlineblojo02184
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Re: Invitro casing layers [Re: Violet]
    #19566242 - 02/14/14 12:54 PM (10 years, 1 month ago)

Got ya.

I've only got second hand plastics cuz I'm still a college kid...

I tried a sgfc with a 2 liter, poor results.
Any write up on the 2 liter you got going?

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OfflinecronicrFacebook
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Re: Invitro casing layers [Re: blojo02184]
    #19566252 - 02/14/14 12:59 PM (10 years, 1 month ago)

don't overthink things, poke some holes...mist when needed:rockon:


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

It doesn't matter what i think of you...all that matters is clean spawn

I'm tired do me a favor

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InvisibleViolet
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Re: Invitro casing layers [Re: cronicr]
    #19566283 - 02/14/14 01:09 PM (10 years, 1 month ago)

There's nothing to write up really.  I cut the 2-liter at the widest point where it just barely fits over the container's threads, and stick it on, with the small lid very loose.


--------------------
Intentionally or not, here in mushcult we are purveyors of love culture and enlightenment movement. Let's try to act like it!

PODS TEK - Growing Invitro with BRF/verm or Grass Seed containers
The simplest, quickest, safest tek!  For beginners, culturers, lazy people, stealth lovers, contam haters, and alternative seekers!
Violet's Teks and Posts

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OfflinecronicrFacebook
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Re: Invitro casing layers [Re: Violet]
    #19566298 - 02/14/14 01:12 PM (10 years, 1 month ago)

:takingnotes:


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

It doesn't matter what i think of you...all that matters is clean spawn

I'm tired do me a favor

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Offlineblojo02184
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Re: Invitro casing layers [Re: cronicr]
    #19566633 - 02/14/14 02:33 PM (10 years, 1 month ago)

Quote:

cronicr said:
:takingnotes:



:lol: ... :takingnotes:

Violet, do you plan on getting bowls similar to cronicr's and testing BE on larger volume subs?

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InvisibleViolet
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Re: Invitro casing layers [Re: blojo02184]
    #19589008 - 02/19/14 08:34 AM (10 years, 1 month ago)

Bottom-watered second flushes coming in.

May sound like a broken record, but again, I can't wait to see how this does with my better cultures.


I love this method. I am kindof thinking now that this is my grow method of choice.
Brown rice, soil casing layer, room for fruiting in the container, no-maintenance except watering which is SO easy and quick. Even if one doesn't purposefully maintain sterility within the container when casing, the somewhat still-air conditions of invitro make spore printing these caps effectively a sterile procedure.  And of course cloning is close to that as well.

May I reiterate:  NO-maintenance! Except watering twice per flush (with good culture) which doesn't even take as much time as one would want to LOOK at them.
The conditions keep themselves perfectly with a loose lid.  It almost does it perfectly invitro for starters, but the casing layer ensures it.
I haven't misted a single one at all.


The only drawback of this method, compared to the "A-tek" as per with the half-size pint containers in fruiting chambers, is that these containers take up double the space in the cooker.  My small AA910 can fit 6 pint containers to sterilize for 40-50min, but of course only 3 of these containers to sterilize for 40-45min.

But the advantage this gives is the complete lack of any kind of fruiting chamber, not to mention the increased reliability and the sterile potential.
As long as one can do a nice clean transfer to inoculate, it's pretty much a Sure-Thing Grow.


Even if one only manages 200% BE from the rice (I often get 300-370% with my good cultures in well-done grows), the ~70g of rice in each container would have yielded ~14g after numerous flushes.  A 2lb bag of rice would yield ~90g in just 6-7 containers.
And if you get 300% BE, 21g per container. 2lb of rice grows 136g.
The culturing-mastery "holy grail" 400% BE, 28g per container.  2lb rice grows 181g.

For me 2lb of rice, prepared and divided into Six quart containers, takes just Two 40-minute cooker runs with my small AA910.


Quote:

blojo02184 said:
Violet, do you plan on getting bowls similar to cronicr's and testing BE on larger volume subs?



Eh, I've been considering trying a few grows to straight coir for seeing if the cultures I'm testing do any differently on bulk substrates since I may be sharing them,  but otherwise No, I have no interest in doing that.


--------------------
Intentionally or not, here in mushcult we are purveyors of love culture and enlightenment movement. Let's try to act like it!

PODS TEK - Growing Invitro with BRF/verm or Grass Seed containers
The simplest, quickest, safest tek!  For beginners, culturers, lazy people, stealth lovers, contam haters, and alternative seekers!
Violet's Teks and Posts

Edited by Violet (02/19/14 08:49 AM)

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OfflineCMOS
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Re: Invitro casing layers [Re: Violet]
    #19589883 - 02/19/14 01:12 PM (10 years, 1 month ago)

I had some going a bit ago, I had a bunch of partially full spawn jars I needed to clean up so I cased em and threw them in my GH to try and get some fruits off them.  Mine were in glass quarts though, and alot took awile to fruit or not fruit at all. (some of the spawn was a couple months old). 

Would like to do some of these to get some good clean prints though.

Quote:

've come to realize that Cubensis likes to fruit away from nutrient density when possible




It was interesting watching one of my jars establishing a mycilium network up the side of the glass then fruiting about a 1/2" away from the casing layer.


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

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OfflineBigGreenMat
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Re: Invitro casing layers [Re: CMOS]
    #19592022 - 02/19/14 08:59 PM (10 years, 1 month ago)

Violet, what varieties have you used?  I tried to get a good print from an in vitro APE grow, but even though it pinned and grew into mushrooms they stopped growing before opening up mushed down.


--------------------
My Growth So Far!

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Offlinecallemann
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Re: In vitro casing layers [Re: BigGreenMat]
    #19703179 - 03/16/14 08:06 AM (10 years, 15 days ago)

Very interesting stuff!

2 questions:

1. How exactly, do you "water" the jars between flushes?

2. How do they get enough FAE with the lid(although slightly unscrewed) on? Wouldn't the air become stagnant in the bottom?

Thanks!

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InvisibleViolet
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Re: In vitro casing layers [Re: callemann]
    #20523794 - 09/05/14 10:32 AM (9 years, 6 months ago)

Just a reminder that this is awesome.


Quote:

callemann said:
Very interesting stuff!

2 questions:

1. How exactly, do you "water" the jars between flushes?

2. How do they get enough FAE with the lid(although slightly unscrewed) on? Wouldn't the air become stagnant in the bottom?

Thanks!




Sorry to have taken so long to respond.  I missed that one.
I'm sure you've figured this out by now, and/or quit caring, but for the sake of the thread:

1.  You just pour in some water!
The cakes will have shrunk away from the sides of the container during/after the first flush.  I add water to that space during/after the first flush (during if the 1st flush yield is nice and big!)
Bottom-watering

2.  Cubensis is quite CO2-tolerant.  Many of the photos depicting "FAE lack" have more than one cause going on, and the CO2 buildup is at problematic levels for reasons that are simply and easily absent in this method.
These cakes are smaller than ANY bulk substrate, and have mycelium only on top of the grains instead of intertwining every tiny piece of substrate matter, meaning there is much less mycelium in proportion to the air content of the containment.
A container 1/4 full of colonized grain will have significantly less CO2 problems than a tub just 1/6 full of a bulk substrate cake.

With the lids cracked, excess CO2 is pushed out of the container.  Although controlled, air mixes freely around the top edge of the container.
You'll see this in the ring of evaporated moisture around the edge of the container when the lid has been cracked a while – also the sign of how loose your lid need be.

Far from being suffocated, Cubensis actually thrives in this environment.  The only mushroom I've seen take to it better is Enoki.


--------------------
Intentionally or not, here in mushcult we are purveyors of love culture and enlightenment movement. Let's try to act like it!

PODS TEK - Growing Invitro with BRF/verm or Grass Seed containers
The simplest, quickest, safest tek!  For beginners, culturers, lazy people, stealth lovers, contam haters, and alternative seekers!
Violet's Teks and Posts

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OfflineGoodyMcLuvin
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Re: In vitro casing layers [Re: Violet]
    #21151972 - 01/20/15 08:03 PM (9 years, 2 months ago)

I'm sorry to bring up this old post, but when I first read "V Tek", I wondered if it had been done in vitro. How would you compare your flushes per half " Mycoquart" compared to uncased half quarts done in a fruiting chamber? And at the chance of sounding like a noon, what's BE?


--------------------
*I graduated from the corner, my respect be my diploma,I went OG for the dollars, Js thought I went to college*

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InvisibleViolet
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Re: In vitro casing layers [Re: GoodyMcLuvin]
    #21152274 - 01/20/15 08:51 PM (9 years, 2 months ago)

No need for apology, for some threads like these it doesn't matter if you bump them.

Biological efficiency is the term for the mathematical measurement of how much yield is produced from a certain amount of substrate before water is added to it.
It's measured by dividing the fresh weight of mushroom harvest by the dry weight of substrate used. And since mushrooms are 9/10 water you can just add a 0, or conversely move the decimal point, to go between calculating with dry and fresh fruits.

Tying that into your yield question -
You mentioned "myco-quart" - but the "myco-quart" is somewhere around 660-750mL.  Half of that is more than I load the quart invitro containers.
Load 1 cup of substrate. The containers have a marker for it too. That, or right at or short of the 250mL mark (I prefer metric).

Since 454g (1lb) of brown rice expands to a little over 6 cups(/containers), about 6.3-6.4 cups estimate, we can divide 454 by 6.3 to have a fairly accurate measurement of how much dry grain is in each container. Comes out to ~70-74g rice per container.

Depending on the strength of the culture for the sub and method and if you water adequately, you can typically get anywhere from 200-300%BE. From 72g rice, that's about 14 to 21 dry grams. With multi-spore or so-so cultures you may not get 14, and you are unlikely to get 21 without looking for top-notch cultures such as with my culturing tech.
Also, invitro MIGHT have a slightly shorter yield but I have not confirmed this.


--------------------
Intentionally or not, here in mushcult we are purveyors of love culture and enlightenment movement. Let's try to act like it!

PODS TEK - Growing Invitro with BRF/verm or Grass Seed containers
The simplest, quickest, safest tek!  For beginners, culturers, lazy people, stealth lovers, contam haters, and alternative seekers!
Violet's Teks and Posts

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