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TV Guide to Colonizing a Quart Jar in Three Days


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#1 TVCasualty

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Posted 26 December 2007 - 01:21 AM

This is a cheap and relatively easy way I've come up with to use BRF half-pints to inoculate grain by turning the colonized BRF into a liquid slurry that is then poured into quart jars of whole grain. I had seen the blade assemblies of some brands of household blenders used in this context before with colonized petri dishes, and I tried it as an alternative to the very expensive autoclavable blenders sold for this purpose. I didn't expect the incredible results I got from using a BRF jar instead of a petri dish as the source of mycelium, and after some refinements to the initial design I've come up with one that provides consistent results, is extremely durable, simple to make, and cheap.Although the procedures themselves are fairly simple, this probably qualifies as an intermediate or advanced tek. It's not a good choice for a first grow and I've made some assumptions in these instructions, namely that the reader has grown mushrooms before and is familiar and experienced with the PF Tek, growing on whole grain, and creating proper fruiting conditions, none of which are covered here.Here's a summary of how it works:

  • Choose a good looking half-pint BRF jar colonized with the species/strain of your choice. I recommend a lighter, bare-bones BRF recipe that allows for the quickest colonization times (I use about 1/4 cup of rice flour and a pinch of corn meal per half pint). I usually only fill the jars slightly over halfway with substrate so the dry vermiculite contaminant barrier is very thick. Think of it as a three dimensional petri dish.
  • In front of a flow hood, dump the cake out of the jar and cut off the upper portion just in case it harbors a contaminant.
  • Drop chunk of colonized BRF cake into sterilized mason jar containing about 200 ml of water, using a blender blade assembly as the lid (tighten the band firmly!).
  • Blend until it's a uniform slurry, usually takes several 3-5 second bursts.
  • Switch blades for lid with dispenser nozzle.
  • Pour slurry into grain jars.
  • Shake jars as vigorously as you can for about 20 seconds each (a long time, in other words).
  • When fully colonized (95% colonized works too), shake them again the night before spawning to bulk or casing the grain. The following morning they should show significant recovery if the temps are correct, otherwise suspect a bad batch.

There are two ways to use this as described: Colonizing jars of grain extremely quickly or inoculating a large number of jars from a very small amount of spawn. So far I've only been interested in setting speed records for quart jars, and my fastest time so far was a quart half-filled with 100% cracked corn that fully colonized in 56 hours (though I don't use cracked corn anymore for various reasons).Materials Required:

  • Regular mouth mason jar (pint size)
  • Blade assembly (including gasket) that fits regular mouth mason jars.
  • Autoclavable plastic lids or regular metal lids, either will work but I prefer the plastic ones (and they are worth every penny)
  • 1/2:; hose barb X 1/2" FIP threaded brass adapter
  • 1/2" MIP thread X 3/8" FIP thread brass hex bushing
  • 1-1/8"; O.D. X 23/32" I.D. rubber grommet
  • Teflon tape
  • Silicone sealant (any type that works for an airport)
  • Double-sided tape
  • Aluminum foil

Tools:

  • Drill
  • Two crescent wrenches that can open to 7/8"
  • Small bit for pilot hole (~1/8"; is fine)
  • 5/8" countersink bit (this was $9 at the Big Box store, but worth it)
  • ~ 3/4" tapered abrasive grinding bit
  • hammer
  • nail
  • scrap wood as backer for drilling

The Blender Blade Assembly:
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This is the easiest part. Get a blender with a blade assembly that is the same diameter as a regular mouth mason jar (approx. 2.5 inches or 65 mm). Oster brand blenders have several models of this size, and if you already have a blender or want to make several mason jar blenders, the cheapest source for just the blades and gasket is here ($6.99 each).By itself, the gasket tends to get tangled up in the blades, which was a problem I tried numerous ways to solve.

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The simplest and most effective solution was just sticking the gasket on with some double-sided tape. Use four pieces of tape to stick the gasket to the base, then trim off any extra tape around the outer edge. It doesn't hold very tightly, but it's only there to add just enough adhesion to avoid the entanglement problem. DO NOT use silicone or anything else to attach the metal band to the blade assembly. I discovered the hard way that the band must be loose or you cannot tighten the lid enough to prevent leaking. You can also RTV silicone the gasket to the lid on the lid side. This seems to be the best solution.

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The Dispenser Nozzle:Here's a photo of the parts needed for the dispenser nozzle. I will show one being made with both a metal and a plastic lid.

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These are the tools, and I've already marked the center of the lid. The hammer and nail are just for punching a little dent where you want the hole so the drill bit doesn't wander across the lid.

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Drill out a pilot hole, using some scrap wood as a backer.

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With the 5/8"; countersink bit, expand the hole until the bit passes all the way through. This can be tricky at first, so use light pressure and medium RPM to get the hang of it. Be careful not to crimp the lid anywhere or it won't work well and makes a mess later. The bit works well in wood, metal, or plastic.

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Now expand the hole further with the abrasive bit. This also takes a little practice to get good at, and I suggest holding the lid in one hand and rolling your wrist so no part of the bit gets a rut dug into it by the metal. Hold the drill firmly at max RPM with your other hand. It's not hard to make a nearly perfect hole if you're patient, and have a few practice sessions. This bit also works on plastic (watch out for it clogging up with plastic, a stiff brush can clean it off).

 

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This is the step that requires the most patience. Still using the abrasive bit, keep shaving off material a little at a time and then test the grommet to see if it fits. It should be easy to attach it to either type of lid, and it's ok if the hole gets a little too big. When the grommet spins freely in the hole, it's big enough. Ideally, to fit the threaded brass fitting through the grommet, you should have to screw it in. The brass threads will cut into the rubber as you twist, and so long as it'll fit through far enough to attach the nozzle it's fine.


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This is the view from the bottom and side. You want the threads oriented this way or it won't work.

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Now wrap the threads with Teflon tape. This makes it much easier to take it apart later if necessary.

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Using the wrenches, tighten the barb fitting (the nozzle) onto the taped threads. As the barb screws on, the grommet will compress a little, so you can tighten it further than it appears and the fittings can handle a lot of force, so don't be shy when you tighten it. Watch your knuckles!




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Clean the lids really well, then seal the fittings and grommet to the lid with silicone (seal both sides of lid). Let cure at least 24 hours.



The finished dispenser lids.

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Edited by eatyualive, 31 August 2014 - 12:21 PM.
fixing format, old code update pics


#2 TVCasualty

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Posted 26 December 2007 - 01:22 AM

Now it's time to get everything ready for the pressure cooker. Gather the dispenser lids and the blender lids, along with a mason jar, a kitchen knife or spoon, and aluminum foil.

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http://mycotopia.net...=1&d=1198650755

You'll need to cut two pieces for each dispensing lid, one that's about 4" X 4" and one that's 12" X 12" (the pieces in the pic are not the right size, but illustrate the idea).

http://mycotopia.net...=1&d=1198650755


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Take the small piece and make a cap for the end of the nozzle. The easiest way is to take a dowel or piece of tubing (5/8" diameter works perfect) and wrap the foil around it tightly, twisting in one direction and folding one end over. Remove it from the dowel or tube and slide it onto the nozzle. Be careful not to crimp the foil into the barbs of the nozzle or you won't be able to easily (and therefore cleanly) remove it later.
http://mycotopia.net...=1&d=1198650755

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Next place lids in the center of the larger piece of foil.
http://mycotopia.net...=1&d=1198650755

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It's important that you wrap them carefully; first gather up the corners.

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http://mycotopia.net...=1&d=1198650755

Next, wrap the folded-up corners by gently twisting them around the nozzle, then bend the top down as in the picture and your lid is ready for the cooker. Being careful here makes things a lot easier later, so start over if it gets messed up.

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http://mycotopia.net...=1&d=1198650755

To prepare the blender jars, cut a square of foil about 6" X 6", fill the jar with 200 ml of water, and place a blender blade w/ gasket on the jar.

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http://mycotopia.net...=1&d=1198650367

Secure the blade with the metal band, and cover w/ foil.
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http://mycotopia.net...=1&d=1198650367

Take a knife or spoon and wrap it with foil.

 

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http://mycotopia.net...=1&d=1198650367 http://mycotopia.net...=1&d=1198650367

Now comes a part that's a little tricky to describe but simple in practice. Cut a 6" X 6" piece of foil for each blender jar. Wrap it/them in an outer envelope that's also made of foil so the pieces remain sterile after being removed from the cooker.
(pic shows open envelope full of foil squares, fold flap over and crimp lightly to seal).

 

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http://mycotopia.net...=1&d=1198650367

At this point, I place all the parts into a stainless steel pot that fits into my pressure cooker, and I cover the pot with yet another piece of foil that I tightly crimp around the rim. I put the pot in the cooker and PC for 30 minutes at 15 psi, which is probably overkill but too much is better than not enough, right? I can usually PC about 6 complete "Blender Teks" in an AA 921 (including all jars w/ blades, knives, lids, and foil squares). Making extras is a lifesaver for the inevitable screw ups like dropping something on the floor or whatever.

While the tools are cooking, clean your lab space and turn on the flow hood. It's best to let it run at least 30 minutes after turning it on before doing anything in front of it. Let the pot cool off in front of the hood (with foil cover still on).

Lay out the gear in front of the flow hood, and pick a colonized half pint that looks good. With the lid still on and while wearing latex gloves, wipe the outside of the half pint with a paper towel soaked in alcohol to clean off any contaminants that might be on it.
http://mycotopia.net...=1&d=1198650367

 

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The next step is something I do to be extra cautious, and in most cases is probably not necessary, although I do it every time. Remove the lid from the half pint and dump the dry verm into something. With your gloved index finger, scrape/wipe as much loose verm out of the jar as possible. Then take an alcohol-soaked paper towel and wipe the glass inside the jar from the rim down to the substrate so it's squeaky clean (we want to eliminate any nasties that might have ended up there).

Taking the foil square off the top of the blender jar, place it on your workbench so the side facing the jar is now facing up. Smack the BRF jar onto the foil square so the cake slides down.

 

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http://mycotopia.net...=1&d=1198650367


The following step is one that should be performed as quickly as possible. I suggest practicing the motions first until you're comfortable with them.

Unwrap the knife or spoon, tilt the jar over so the cake begins to slide out, then cut about a quarter or so of the top of the cake off and let it fall aside, being careful not to let the larger chunk fall out of the jar. Set down the knife and with a smooth, quick motion lift the half pint up to the blender jar, lift the blade/gasket off the blender, dump the BRF chunk into the blender, then quickly replace the blade assembly. Discard the knife, half pint, foil piece, and chunk of BRF substrate. Clean up any spilled verm or BRF.

 

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http://mycotopia.net...=1&d=1198650367


Open the foil envelope and get a 6" X 6" piece out. Place it over the blade assembly and secure it with the band. Tighten the band firmly. Tear off enough foil to expose the part that attaches to the drive shaft of the blender (see picture). The foil works like an additional gasket to prevent spills in case something doesn't seal properly. I didn't do this at first, and it's often unnecessary, but they never leak when I use the foil.

 

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http://mycotopia.net...=1&d=1198650367

Take a clean blender base (one dedicated to the lab is ideal, and this model is cheap) and wipe it down with alcohol, letting all the alcohol dry off before plugging it in. Put the blender jar on the base, which you'll notice doesn't hold it very well. I found the best way to blend it is to hold the jar with one hand and the base with the other. My hands are big enough to hold the base and press the buttons at the same time, so I hold it all at about a 45 degree angle (the chunk floats so tilting helps get it to hit the blades) and press the Pulse button for 5 second bursts (on "High") and slightly shaking the whole thing while it blends. Let it settle down and check to see if there's any big chunks still floating. Keep blending until the chunks are gone.

 

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http://mycotopia.net...=1&d=1198650367

Your slurry should look like this.

 

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http://mycotopia.net...=1&d=1198651080

Next, remove the band and foil from the jar. Unwrap the dispenser nozzle from the foil (hope you wrapped it carefully!) being careful not to lift the base off the foil. I press down on the brass fitting while unwrapping so the "inside" of the lid is not exposed to the air.

 

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http://mycotopia.net...=1&d=1198651080

In one smooth, quick motion remove the blade assembly and replace it with the dispenser nozzle and tighten. If using the metal version, secure the lid with a band (the one you just took off the blender jar is fine).

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http://mycotopia.net...=1&d=1198651080 http://mycotopia.net...=1&d=1198651080

Arrange 10 quarts of sterilized grain in front of the flow hood. Loosen the lids of the quarts.

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http://mycotopia.net...=1&d=1198651080

I was unable to photograph the following step, since I did it by myself. It really requires a video to show it right anyway. What you want to do is gently shake up the slurry in the jar, remove the foil cap from the nozzle (I hope you didn't crimp it on too tight!), set the cap down, then quickly and methodically open a quart jar and roll your wrist slightly as your pour a shot of slurry into it, then replace the lid of the quart and move to the next. Tighten the quarts after inoculating all 10. Shake them at least 15-20 seconds each. Each jar of blended slurry can inoculate 20 quarts if going for full colonization in under 4 days or up to 50 if trying to stretch the spawn as far as possible. Practice with water and an empty quart jar. The biggest problems here include spewing slurry on the jar threads or all over the outside of the jar (or both), holding the exposed nozzle outside the flow hood air stream (easy on small hoods like mine), or getting slurry on your filter. The stuff tends to have a mind of its own sometimes, so be patient and practice makes mostly perfect.

You can calibrate your dispenser with a small measuring cup. Put 200 mL of water into a pint jar and cover with the dispenser lid, then turn it upside-down over the measuring cup until it stops flowing and take note of how much comes out per shot. Then try two shots, and so on until you know exactly how many shakes or pours it takes to inoculate at the rate you desire. I measured almost exactly 10 mL per shot, so 2 shots per quart gives me 20 quarts. I realize the chunk of BRF cake will add to the volume in the blender jar, but due to the thickness of the brass fitting holding the nozzle you won't be able to use all the slurry anyhow. These things become obvious once you make one and play with it awhile, and another advantage of practice/experience is that each quart will get exactly the same amount of inoculum so they'll all finish simultaneously.

Here are typical results I get with this setup (I took pics when I could):

Just after inoculating, holding the temp at about 80 degrees F (26 degrees C).

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http://mycotopia.net...=1&d=1198651080

The same jars 54 hours later (2 days, 6 hours), then again at 68.5 hours (early morning of 3rd day).

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http://mycotopia.net...=1&d=1198651080 http://mycotopia.net...=1&d=1198651080


It was ready by the morning of the 3rd day (but I wasn't), and at 77 hours from inoculation it looked like this and was becoming difficult to shake.

post-5901-138185699542.jpg

http://mycotopia.net...=1&d=1198651080

Well, there you have it! This is the Tek I'm most proud of, since I get such great results from so little input, and I can see a huge potential for expanding on it and improving it; I'm counting on the mad scientists in the OMC to do just that. My goal with it is to use autoclavable tubing and some kind of pressure system to allow using this without ever exposing substrate/spawn to air, possibly eliminating the flow hood or glove box completely (and yet still being able to do whole grain). I believe it will require modifying the mason jar (I'm drilling holes in them these days).

One more thing: This goes so fast that even a mildly contaminated culture can be used, which can be good or really bad. I discovered this accidentally after having a batch of 20 quarts contaminate almost entirely all at once (inside the substrate), but it only happened after I'd poured the quarts into two trays, cased them, and harvested a flush! Going backwards, I traced the contaminant to my spore syringe, and half pints I didn't blend (which were fruited in the normal PF Tek manner) also contaminated (it was not very virulent, but there nonetheless). If I'd spawned those quarts to a bulk substrate, it's likely I'd have lost the whole thing. That's why I make an extra-large dry verm barrier in my half pints and cut off the top of the cake before blending. A clean culture in the half pints is essential for success.

This is the first in a series of threads that will eventually detail my methods from start to finish, as time permits (and not necessarily in any order). Barring contamination, I can easily produce 20-30 quarts of colonized grain from 2 cc's of spores in 18 days if I'm on top of it, meaning my quarts are cooling off from the PC the morning the half pints are finishing up (I rarely am that organized, but I've done it a few times). It takes 2 weeks for the BRF half-pint jars to colonize from spores, 4 days after that for the quarts to be finished, and if I'm casing them I get my first harvest about 2 weeks after that. At that point I can spawn to bulk or case them, and if casing I can get my first harvest in 32 days, though 38-40 days is more the norm for my lazy self. A lot of magic comes from 20 quarts, but precise yields are dependent on many factors, some of which I will deal with in future posts.

Oh, and all my talk of casing might be obsolete in a month or two. I'm trying to combine Faht's late casing tek with a crazy idea of my own, and I'll post results as they come in.

Finally, I'd like to give a big thank-you to Mycotopia :bow:. The ideas and knowledge I used to put this together came from reading the archives and threads of the many brilliant and experienced people that hang out here. Keep up the good work!

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Edited by eatyualive, 17 August 2014 - 11:28 AM.

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#3 donpuro

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Posted 26 December 2007 - 08:34 AM

excellent write-up... The colonization times in this thread are :bow:

#4 fedshtkpndrk

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Posted 26 December 2007 - 10:05 AM

This is pure gold, TV :bow:

-feds
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#5 TVCasualty

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Posted 26 December 2007 - 12:57 PM

Thanks!

I'll probably be adding a few things as I think of them, and one I'll add right now is that at first, I thought every single jar I inoculated with this method had contaminated. It turns out that as the mycelium recovers and begins to grow into the grain, it looks exactly like grayish cobweb and nothing like bright white mycelium. I was disappointed and thought I'd lost everything, but the next day it was much more dense and bright white, with rhizomorphic strands beginning to form, so I relaxed.

Also, this boosts the temps in the jar very quickly, so I try to keep the room at around 80 for the first day or two, then bump it down a notch to 76-78 degrees after that.

#6 camMyco

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Posted 26 December 2007 - 01:01 PM

I have never learned so much from a TV_Guide!

#7 tecnikal

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Posted 26 December 2007 - 01:06 PM

thats awesome! I definitely see that in my future

#8 eatyualive

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Posted 26 December 2007 - 01:22 PM

osterizer blenders fit a 1/2 pint perfectly on the end. tis goo d for liquid cultures.

and tv, very nice and thorough. me likes the pictures. very explanatory.
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#9 ShroomGuerilla

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Posted 26 December 2007 - 03:03 PM

MMmmmm Milkshake....
:pirate:

#10 Nzo

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Posted 26 December 2007 - 03:51 PM

lol milkshake..

i dont think i've ever seen this tek before. awesome!!
im still reading it cause im kinda confused (im sober lol)
but creative!

#11 WHITE RABBIT

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Posted 26 December 2007 - 04:18 PM

Thats a pretty cool trick TV! Glad you showed us!:rasta:

#12 SporeCrazy

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Posted 26 December 2007 - 05:15 PM

Excellent write up... I've been looking into a fast effective way to do some bulk grows and this method looks killer! You'll have to let us know what kind of results you achieved from those jars! :amazed::cool:

#13 fahtster

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Posted 26 December 2007 - 09:47 PM

pretty sick chubs! :) nicely done. :thumbup:

fahtster

#14 Hippie3

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Posted 26 December 2007 - 09:58 PM

:bow:
archive material

#15 OG Royal Grower

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Posted 27 December 2007 - 01:00 AM

mad props

#16 the_chosen_one

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Posted 27 December 2007 - 01:56 AM

WOW bro! you weren't kidding! this is some piece of work! :eusa_clap

#17 Lazlo

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Posted 27 December 2007 - 08:36 AM

Nice!

#18 TVCasualty

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Posted 27 December 2007 - 12:25 PM

Thanks for the good reviews! So what do y'all think we ought to call this thing?

When you see the parts to this sitting in front of you it doesn't look very complicated, and really it's just making a myco-smoothie in a way that involves a lot of aluminum foil, but writing it up with all the relevant details (and without it getting ridiculously huge) was harder than doing it! :puke: I was trying to give enough detail to allow someone with minimal familiarity with tools to make it, but it does require some fabrication skills, so I'd been toying with the idea of making lots of the dispenser lids and maybe selling it as a small kit.

I think it needs further refinement to be a retail item, and when time permits I'm going to make a lid with two medium-size nozzles instead of the one large, and use autoclavable silicone tubing to eliminate the open-air pouring by pushing filtered air into one nozzle to force the slurry out of the other. With a large syringe (no needle) as a source of compressed air, I can push precise amounts of the slurry into jars by pressing the plunger of the syringe, or at least that's the plan. With a large-bore needle on the end of the tubing and a silicone port on the jar lid, I could inoculate jars without opening them.

Also, drilling a hole in the side of a jar and covering with silicone works well for an airport and frees the lid area for another purpose, like blender blades.

This setup works well for making isolates, and I've used almost an entire stem as the source material, which I can do thanks to the sharp blades (just mix up the water with your favorite LC formula instead of plain). I peeled the outer layer off and dipped it into a dish of peroxide before dropping it into the blender jar, and the blades turned it to mush. It recovered and grew fast thanks to using the whole stem instead of just a little chunk, and I left the blades attached until it was ready to blend again just before using it to inoculate the next step. For air exchange, I had an airport stuck in the hole I'd drilled in the side of the jar. You can draw the isolate back up through the port or switch the blades for the dispenser lid. When making isolates, it's good to have a few redundant blender jars available as cloning is more challenging than blending the BRF cake.


One thing that's been bugging me for a long time is while I'm certain I'm not the guy who came up with using these blades on mason jars, I can't seem to remember or find the place I'd seen it described before (years ago), and so far I have not read of anyone around here using them. So, what's up with that? Anyone else out there know of someone using blender blades on jars for home mycology?

Oh, before I forget, check out this interesting math: A case of 12 half pints of colonized BRF (using 24 cc's of spores) can provide you with 240 colonized quart jars within three weeks of shooting the spores (that's a theoretical maximum, anyway). The 12 blender setups would cost about $200, and I included the cost of a cheap drill! Compare that to a blender designed for lab use; a SINGLE 250 mL Eberbach blender can cost $250, then add the required $180 base and you can inoculate about 20 quarts at a time for just under $500 including shipping. It'd cost almost $750 to get the 1000 mL model+base, and that's only good for about 100 quarts at a time. So, being able to inoculate 240 quarts for $200 instead of 20 quarts for $500 seems like a good deal, and makes me think this is an option for a commercial edibles grower on a low budget or just starting out (and you get to keep the drill!).
  • tailsmcsnails likes this

#19 joystik

joystik

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Posted 27 December 2007 - 07:49 PM

My respects to you. Excellent writeup!! Invaluable information.

#20 GordianHyphae

GordianHyphae

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Posted 07 January 2008 - 10:57 PM

First of all, :bow:! Fantastic idea, implementation and write up! I am much inspired, and it turns out, I've already got an osterizer blender, the base of which screws perfectly onto a regular mouth canning jar. I'm intrigued about your pressure dispensing idea, and it occurred to me that once you plunge the air syringe to pour a shot, you'd need to pull the tubing off and suck more air into the syringe to push the next shot out. This air would need to be filtered or else it's a contamination vector. What if, instead of air, you used a mini co2 injector, like this one. it costs $30, with replacement cartridges at a buck a piece. each cartridge holds six liters of pressurized co2, which is bound to be pretty damn sterile. It says on the site that the trigger mechanism allows a measured injection. It seems like it would be pretty easy to rig this up to a jar. just an idea i thought you might be able to work with. they also sell injectors like this at bike shops for inflating flat tires. i'll be tinkering with your ideas over the next couple of weeks, and i'll let you know if i figure anything cool out. Thanks again TVC!




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