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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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).



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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.

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Next place lids in the center of the larger piece of foil.

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





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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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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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Secure the blade with the metal band, and cover w/ foil.



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Take a knife or spoon and wrap it with foil.

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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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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.

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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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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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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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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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Your slurry should look like this.



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



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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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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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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.

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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.