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Maximized Invitro Bag Method!

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#1 morthos



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Posted 18 May 2005 - 03:19 PM

Afternoon folks! After the successes of my Invitro rez-effect oven bags, I'm looking at ways of maximizing the method. After some PM discussions with Uncle Fester, I feel we've got some ideas that have value.

I'd mentioned before that I think a 6L mass can produce more that what it has for me so far (tho it's been good already:)). We all know that a PF jar gives limited fruits based on the resource/demand ratio, where a lack of resources makes for a bottleneck production level for that mass. I figured that such as thick mass should be able to produce more; in other words it's current shape which has some parts about 6" thick vertically has exceeded the resource/demand bottleneck and has more mass or thickness that can be used. The logical thing to do is to increase the demand on the same mass to get a higher yield. My theory to do this with is to increase surface area.

While some have brought up the idea of suspending the mass to encourage fruiting underneath, I don't see a way of doing this well with my setup; I'm using plastic bags & metal shelving units...even if I could suspend the masses, that would force them to go into a bowl or sphere shape - a sphere being the minimum surface area shape.

My method will be to form the mass into thinner, rectangular shapes. Being further from a sphere shape it can only have greater surface area, and the mass will only be 3~4" thick - hopefully closer to a thickness that can be more efficiently used for fruiting. I'm aiming to get a thickness that meets or comes just under the resource/demand surface area bottleneck. Unused resources=wasted resources. Excess surface area=insufficient resources (tho no wasted resources). Maximum use of resources=maximum yield=best use of my time & effort! :)

My first thought was to do this within the existing oven bags, and see what yield improvement there was. I'd only made them the way I had because I knew I could close the bags easily in that shape! But why stop there? Why not take up the whole 11"x27.5" shelf space with a mass? Maximize the use of given space! Using a clear plastic tray, 4" deep, for support, and a clear garbage bag with 2~4 breathing patches? These bags should come sterile, so I should be able to attach the breathing patches without PC'ing given I sanitize the area first. As this couldn't fit in my glovebox, this method would demand I use all-colonized substrate...spawning not advisable. I'd still PC my moistened verm just to be safe. The garbage bag would be tied shut at the end, while any excess plastic would be folded to one side & held there by, say, office paper clamps, or clips, or whatever, until fruiting demanded they be removed to there's enough space to grow inside the bag.

The total volume of mix in the bag would be determined by filling a test bag with plain verm and forming it into shape in the plastic tray (verm returned to source when test finished, eh!). Knowing this volume, one would prepare half that in colonized wbs, and half in PC'd wetted verm. After all that spawn & verm is prepared, one could empty it all into the prepared garbage bag in sanitized open air, close, then form into shape in the plastic tray, clamp the excess plastic, and come back a week or two later for fruiting. During fruiting, the clamps would be removed or scaled back to give easy growing space.

It'll take me close to 3 weeks to generate the colonized wbs for this; two weeks for spores to colonize a large # of 500ml wbs jars, then another week to spawn them in my glovebox & colonize legions of others.

My main Q to you folks is: what to you think the ideal depth of mix is to be used for fruiting? I'm thinking in the area of 3~4".

What do you think?

#2 Hippie3



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Posted 22 May 2005 - 06:40 PM

fill the 4" tray to the top
better air exchange that way

#3 morthos



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Posted 22 May 2005 - 10:38 PM

Cool, thanks. Hopefully I'll have this running b4 the 3-week mark; I've got 21 jars inoculated with spores or clone matter. Some will be spawned, others used asap.

This will be fun, hopefully epic!:)

#4 morthos



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Posted 02 June 2005 - 10:44 AM

My legions gather for the challenge! That's three dozen freshly spawned jars on the bottom. Two more dozen will be made! One week, and it's time to mix. Stick around folks, soon you will see my ultimate method!

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#5 cheeseball



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Posted 02 June 2005 - 12:17 PM

Morthos, you think you might do a step by step for these oven bags? where did you purchase them? Not to keen on the idea of invitro like that, but I can not argue with your results, so I may just have to go the other side. I'm not gonna quit casings entirely, unless the bags make me :) ;)

#6 morthos



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Posted 03 June 2005 - 12:34 AM

Cheeseball, I'll be honored to do a proper write-up of my OBIRE tek including step-by-step action shots of everything, particularly patch & bag building. To do properly will take me some time, but I'll try to get it here soon. In the meantime you can find all the info under "Biblical Invitro Oven Bag Rez-Effect idea" (note I've rearranged the words for the OBIRE acronym, just so it handles better :))

This thread is regarding the maximum version of OBIRE mind you; it will be roughly 11"x27"x4" in a clear garbage bag with multiple breathing patches. The length and width have been determined by my shelving unit. The total volume I've yet to determine, but that will be pretty easy. I won't be using all my spawn on one bag.

I buy oven bags at the local grocery stores. I'll send a bag to your door to convince you; you'll find he makes an labor & space efficient argument! :)

#7 Guest_pissybee_*

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Posted 03 June 2005 - 07:22 AM

So, basically, you are doing invitro bags in the shape of rectangle dishes? Like invitro rez-effect casings, correct?

#8 morthos



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Posted 03 June 2005 - 09:54 AM

Yes. I'm doing the rectangle shape with both this overgrown version and the oven bag size. The former due to the shape of the shelf, the latter so that there's more surface area than how I first did it; at first the oven bags were a big mass at the end of the bag with the middle 6" thick - I don't think the center resouces were being used. I'm now shooting for a maximum 4" thickness regardless of bag volume to get the most out of the material.

The rest of my bags will be 50/50 wbs/verm, yes. I won't be bothering doing popcorn given the $ difference...I'm getting wbs for less than $1/kg!

#9 morthos



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Posted 10 June 2005 - 11:40 PM

Here we go folks!

I've prepared my first max-Obire bag, coming in at a 14L volume of wbs/verm! That's 17 500ml wbs jars estimated at 400ml material each, plus 14 jars of PC'd wet verm. The garbage bag used for this can't be PC'd without melting it. Oops.

I started by making a box to support the bag for moving. Pic#1 is that, 3" high, 11" deep and just over 2' wide. Just cardboard & duct tape. As such it supported the weight well.

Next I planned how to lay the bag in, and where to attach my patented :) breathing patches. (pic#2) The idea is to fill the box, while having extra plastic available vertically later. Until pinning, there will be a twist tie close to the mass, and another at the top of the bag. The first will be removed to give space for growing.

I sanitized the air and dumped the jars of verm & crumbled the wbs into the modified garbage bag. To mix I tied shut the bag, then put it into a standard, stronger bag so I could mix the bag without fear of tearing the bag. These clear bags don't feel very strong. I then fit the bag into the box and worked the mass around to fill the box as best as possible (pic#3). Here I realized I could fit 15 or 16L in just as easily. Properly fit, the extra air was squeezed out and the twist ties placed in position. (pic#4)

Finally I placed the box on my growing rack. (pic#5) The duct tape on the corners was cut to expose more surface area and maximize pinning. When it needs to be moved, I'll just bring the flaps up and re-tape or support the bottom with something else. (the OBIRE's underneath are the re-mixed bags described in the 'peroxide use with colonized matter' thread, and are exceeding expectations)

This mass is more than double the 6L obire's before it, and I'm hoping to get a proportional increase in yield. Other than that I'm using my space more efficiently. The next two weeks will be very interesting!

I'll be making a few more of these in the days/weeks to come...just a matter of spawning & PC'ing verm! Victory can be so easy when you get systematic...! :)

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#10 Hippie3



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Posted 11 June 2005 - 05:04 AM

good luck

#11 mrco0o



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Posted 22 March 2006 - 11:51 AM

Hey Im new to the board but not completely new to growing and trying to get back into it. I am fascinated by Morthos's oven bag bulk invitro tek because that would definatley work in the situation I am in(TINY ROOM). I had a couple questions though and wanted to see if anyone would be able to help out. My plan as of now would be...

- Prepare several BRF jars, sterlize, and use as the bulk substrate in the oven bag.
- Prepare 1 or 2 BRF jars, innoculate, and wait to 100% colonization. Then add this to the bulk BRF and mix up.
- Grow invitro from there on.

I skipped the basic steps like preparing filter patches and stuff just so you dont have to read it that just an overview.

My questions are...

- Morthos recommended a 2:1:1 ratio for food/verm/coir but this was using WBS. What kind of ratio would I be looking at for BRF since that is prepared with verm? Also, I cannot find coir anywhere, would there be a good subsitute for it?

- Ive read every invitro tek I could find and still a little confused on what they mean by introduce to light from the beginning. Just introduce a couple hours of light from the beginning or 24 hours of light the whole way?

- And finally what kind of temperatures would I be looking at for this tek? Should I incubate at 80-83 F then drop it down towards fruiting time or keep some sort of uniform temp throughout the whole invitro process?

Any help is appreciated! :cacti:

p.s. - im not really interesting in using WBS or anything of the sort as a substrate. Im doing that now for a few jars and I have more birdseed on my floor then carpet! Just too messy for me lol.

#12 Hippie3



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Posted 22 March 2006 - 11:55 AM

just omit the coir and use ratio 2:1:1 verm:water:seed
grind the seed like brf.
Just introduce a couple hours of light from the beginning
incubate at 80F then drop it down to 75F towards fruiting time

#13 mrco0o



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Posted 22 March 2006 - 01:12 PM

thank you kind sir.

#14 viraljimmy



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Posted 22 March 2006 - 05:14 PM

So... Do you think bags will produce
more than jars in the same space?
Just curious.

#15 morthos



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Posted 23 March 2006 - 01:18 AM

Good day mrco0o! Nice to see someone reading my work!

In order of your post:

- Hippie3 got that one

- I left my Obire bags out all day, and natural light only. I had them on a metal rack with a plain sheet over it. I've never tried them in a closet...enough light gets through a cheap white sheet, and keeps them out of sight. Unless you have to grow in a sealed space, I recommend that you avoid timed unnatural lighting. I've never tried, mind you.

I like to encourage pinning asap, so I expose them right from mixing time.

- I didn't use controlled temp varied from 74~80F.

As for what I think of your method, it is functional in principle, but I have some suggestions/Q's...

I found that brf is a dense, hard subtrate to work with and not ideal for bulk. I used wbs as it is much easier to work with, cheap, doesn't need an eternity of grinding in a coffee grinder, etc. Between harvests you will have to crumble the mass; breaking it up well so water would mix in is labor intensive with brf.

Wbs on the floor..? This can be dealt with!

You would want to be using 500ml jars for the bulk substrate, of course. Again, a brf 250ml jar takes as much as four weeks to colonize, where properly used wbs takes two...

viraljimmy, I do feel it produces more than jars, in the way that the space occupied on a shelf will produce much more becuase more of that space is occupied by growing matter. I don't know if it produces more for the same weight; I have never considered it. In a spot you could place twelve 250ml jars(3L total substrate) you can place a bag (6L), and it is less labor intensive. From a 6L bag I was getting an average of 2.5~2.75 dry oz from the first harvest.

I actually have most of my tek written, but it's not complete. Let me know if you want what there is. I'm moving, so I won't be able to finish it for a few weeks.

I look forward to dicussing this mrco0o!

#16 hogwild



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Posted 23 March 2006 - 02:09 AM

4 weeks to colonize a 250ml brf seems a little excessive.

#17 Hippie3



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Posted 23 March 2006 - 07:41 AM

Let me know if you want what there is.

yes plz, sounds like experience has made a few changes in it.

#18 morthos



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Posted 23 March 2006 - 09:39 AM

Morthos’s Obire tekOven Bag Invitro Rez-Effect

This tek was intended to mimic a PF cake grown invitro on a larger scale. Instead of 250ml jars, oven bags of 6 L are made. Time from mixing/rebagging to harvest is 14~18 days with a 6L wbs/verm mix and yields 2~3 dry ounces on the first flush, with declining-yield multiple flushes.

This is a rez-effect method, as is the PF tek. In this case the ratio of wetted verm to prepped wbs is 1:1. The volume & shape used in this tek is 6L based on the idea of a large resource base with good surface area, with 18”x22” oven bags as the cheap and plentiful tool. I did not want to be ordering a lot of spawnbags so I went with what I found at the supermarket.

The general method to make a bag is as follows:

1. Prepare & pressure cook numerous 500ml wbs jars.
2. Inoculate above jars. Steps 3 & 4 can be done during germination & colonization.
3. Prep & PC legions of wbs jars.
4. Prepare double-layer tyvek breathing patches. Tape onto oven bags & PC oven bags 30 min.
5. Spawn wbs from colonized jars made in step 1 with jars of step 3.
6. Prep & PC jars of wetted vermiculite. Ratio of verm jars to wbs is 6:8.
7. Take 8 colonized jars from step 5 and 6 verm jars from step 6 and crumble/empty into a bag from step 4. Tie shut, mix bag & place on shelf until harvest time.
8. After harvest, crumble mass into large bowl. Add ½ ~ 2/3 of water lost to flush & mix. Load into new bag, tie shit & place on shelf again. Repeat until flushes are no longer worth the shelf space.

The detailed method is as follows:

1. Prep & PC numerous 500ml wbs jars.

1. Put wbs in pot, cover with water. Boil 10 min.
2. Dump wbs into colander. Use a fine wire mesh in your colander in case some of the grain gets through your colander. Rinse well.
3. Put wbs back in pot & cover with water, enough to cover wbs after soaking & swelling. Boil 5~10min to kill bacteria in water.
4. Cover pot & let it cool off. When it’s no longer hot to the touch, cover the pot with saran-wrap. Refrigerate overnight.
5. Bring pot to a boil. Dump wbs in colander & let sit 45 min.
6. Load jars 4/5 full & PC at least 60 min. The lids have 4 holes around rim, then tyvek, band, and foil. Bands are left slightly loose.
7. Tighten & shake jars 1 hour after PC'ing- when they are cool enough to manage.

2. Inoculate above jars.

Basic principles here…inoculate with a syringe of anything fresh, not spawn material. I feel that since there’s spawning to more jars, then to the verm later, that the mycelium will be stretched enough as it is. Inoculate in whatever method you’re good with…glovebox, flowhood, sanitized air, etc. I use 1~2 ml/jar of spore solution when I do these. Shake the jars before inoculation, inoculate, then shake again. Set on shelf to colonize.

These jars will be used as spawn material for the jars that will be mixed into bags. One jar spawns to enough wbs for one bag. After colonization these spawn jars should be refrigerated until needed; if the jars pin effort will be required at mixing time to remove the fruit, and months can pass before you get through a dozen jars.

3. Prep & PC legions of wbs jars.

Same procedure as step 1. Make 6 jars here for every 1 jar in step 1.

4(a) Prepare double-layer tyvek breathing patches.

I use two layers of tyvek with a metal mesh between so contams can't wick their way across. The size of mesh square cut is the size the breathing patch will be; I make mine about 3"x4". I electrical-tape the mesh to one piece of tyvek then cut around that electrical tape, cutting away a thin width of tape to be sure the air must mass thru the mesh. Then I put that piece upside-down on a larger piece of tyvek and repeat. Refer to pictures.

The metal mesh can be found at hardware stores. Get the finest mesh you can; it needs to be flexible. The electrical tape is unaffected by PC’ing. Still, I like to buy high-temp resistant types for kicks.

4(b) Tape onto oven bags & PC oven bags 30 min.

The patch is then placed tape-up near the bottom of an oven bag, centered, about 3~4" from the end. Your oven bag will fold out, so lay it flat and arrange from there. Don't try to put the patch on the very bottom of the bag on the need! Tape the patch to the bag. Invert the bag, and carefully cut away the plastic that's under the patch. Uninvert (?), fold & squeeze out air, wrap in 3 layers of foil and PC for 30min.

5. Spawn wbs from colonized jars made in step 1 with jars of step 3.

What’s important is that a colonized jar gets mixed evenly with PC’d wbs to make eight jars/3 liters. This can either be done with jars as I do, or one could empty the spawn & PC’d wbs into a spawn bag. Again, use what method you prefer to spawn with…glovebox or flowhood. I’m a glovebox man myself.

1. Place 2 layers of paper towel in glovebox to A)make cleaning up easier B)absorb excess moisture/lysol
2. Place lysol in corner of box. Add in pre-sterilized tools inside sealed lunch baggies including paper towels, exacto-knife, fork & spoon, rubbing-alcohol soaked paper towel, etc.
3. Shake, then place wbs jars in box, still covered with original foil from PC’ing. If a bowl is needed, it is placed upside-down in the box. Add in other required items, wrapped if necessary with foil to protect from Lysol.
4. Add lid to box. Tape down each corner of glovebox with electrical tape to keep good seal during work. Spray lysol in box on the sides and the gloves.
5. Remove arms from glovebox, wait 20 min.
6. Put hands in gloves and get it done. Insides of bowls are to be wiped with the disinfecting towel. In case of fogging from excessive spraying, use dry towels from baggies to absorb it or wipe glass.

Remove foil from all jars & place in a corner of the glovebox. Have the 8 jars arranged so that the empty jar is in the bottom left of the jars, in the upper left of the box (with your bowl in the right and your utensils in the bottom left). Your colonized jar should be on the upper right. The six PC’d jars are also in the upper left of the box.

Begin by crumbling the spawn jar into the bowl with a fork. This will ease process later. Reload the colonized wbs into its jar.

Open a wbs jar and empty some matter into the bowl. Replace this with almost as much from the colonized jar. Reseal. Continue this until all six jars have had spawn added, and the colonized jar & empty jar are filled equal to the others, at roughly a 4/5 level.

Remove jars from box. Shake all jars to distribute spawn evenly. Place on shelf or in box until conolized.

6. Prep & PC jars of wetted vermiculite

This step isn’t mandatory when using colonized wbs to mix a bag. If a person wishes to make the spawning & mixing one event it demands PC’d verm(takes a few more days to harvest).

I do think it is a good idea to PC verm, just for the better odds of good growing. It doesn’t take long anyway.
I don't clean my glovebox; I simply wipe it out. The thing permanently stinks of lysol anyway! I only use bleach for surface sanitizing & bleach dunking. My tools are washed in Sunlight dishsoap water, dried, then wiped with rubbing/isopropyl alcohol before insertion into lunch baggies.

Well, that's what I have so far. Steps 7 & 8 aren't too complicated; please ask if you've any Q's!

#19 morthos



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Posted 23 March 2006 - 09:48 AM

And some pics while we're at it...

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  • Unwrapped 5L harvest.jpg
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  • sm_10 Obires.jpg

#20 Hippie3



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Posted 23 March 2006 - 10:31 AM

one for the vaults
i think
you have perfected
the invitro tek.

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