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Mushrooms, Mycology and Psychedelics > Mushroom Cultivation
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Airtight Incubation Question
    #15750392 -

I'm using the PF Tek. My jars are steralizing right now. I'm just wondering after I inoculate, should I place them somewhere completely AIRTIGHT? I was thinking of putting them into airtight tupperware after I inoculate.

Secondary Question: Just a thought, what do you guys think about a Cork being used as a lid? Would it work? I could just inject the cork... idk
Thanks

Edited by Shroomasaurus (02/02/12 03:54 AM)

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Re: Airtight Incubation Question [Re: Shroomasaurus]
    #15750465 -

During the colonizing stage, the organism will need a small amount of fresh air exchage. As you become accustomed to this forum, you'll see that term (fresh air exchange) abreviated as "FAE".

In fact, the amount of FAE that colonizing jars need is so small that it has it's own term; Gas Exchange, or GE.

Keeping your PF jars in an air tight incubator will deprive the jars of GE, so air tight would be a no-no. The same goes for your cork lid idea. Yes, you could inject through the cork to inoculate, but unless you had a hole drilled in it with a filter patch, it would suffocate the mycelium (myc).

If you prepared your jars correctly, then there should be four holes in the lid and there should be a half inch layer of dry vermiculite (verm) between the substrate and the lid.

Assuming this is all correctly in place, you just inoculate through the holes and set the jars up on a shelf until you begin to see tiny pins growing in the jars.

Although the dry verm layer method is very effective, I've had my concerns about the needle creating a funnel shaped divet in the verm when inoculating. That's why I've constructed these containers.



These containers cost about a dollar for 12 of them at walmart, making them a lot less expensive than glass canning jars. The lid is integrated into the container, making them much more hassle free than jars. They are made of PP5 polymer plastics, which can be pressure cooked (PC'd) many many times without worry of melting or warping. They also don't break when droped and don't rust, a property I've come to loathe about metal jar lids. 

The container schematic pictured above would be used if a dry verm layer was used. This way, the injection sites would be offset 45 degrees from each breather hole, theoretically reducing the chance of contaminates finding their way in through the divets made during injection on conventional PF jars, but I have not yet tested the ones I've made.

I have, however, tested these.



The container schematic pictured above allows for the dry verm layer to be taken out of the equation. Instead of relying on a dry verm layer for air filtration, a square of Tyvek is adhered into place above a 3/8 inch hole. All three of these are currently colonizing well and do not show any signs of contamination.

-EDIT-: You will run into tons of conflicting information about Tyvek and which type to use. There are many types of Tyvek, and many will have varying degrees of success using any one of them. But the best type of tyvek to use is the type they make painters' coveralls out of. But don't go out and buy just any coveralls. They must say Tyvek on the label. I've only been able to find real Tyvek coverall's at Lowe's.



Once you've found some Tyvek coverall's, remove the arms and use them for your glove box or still air box. The rest of the suit will be enough to produce hundreds of filter patches and will open the door to pretty much any type of colonizing container, no matter how unorthodox... As long as you play by the rules of proper sterile technique!


--------------------
The Shaman

Edited by shamanamba (02/02/12 04:55 AM)

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Re: Airtight Incubation Question [Re: shamanamba]
    #15750478 -

Thanks, really informative reply. I'm using a 1/4 pint jar. Here are the steps I took:

1. Pokes 4 holes in the lid.
2. Filled the jar with the proper mixture of verm and brf.
3. Added the dry verm layer.
4. Attached the lid.
5. Wrapped a double layer of tin foil around the lid.
6. Wrapped a second and seperate 1 layer of tin foil around the lid.
7. Steam steralized the jar in a pot.

This is what I plan to do for the inoculation stage:

1. Remove the top layer of tin foil and put it upside down.
2. Whipe the second layer of tin foil down with alcohol swab.
3. inoculate the jar by piercing the tin foil in each of the 4 holes.
4. Whipe the removed top layer of tin foil down with the alcohol swab.
5. Reattatch the top layer of tin foil.
6. Store for incubation

Does this seem right?

Thanks again

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Re: Airtight Incubation Question [Re: Shroomasaurus]
    #15750499 -

Alcohol sanitizes. Sanitary and sterile are not the same thing at all.

The best thing to do would be to flame sterilize the needle.

While the needle is still RED HOT, start your inoculation of the first jar.

While you must flame sterilize between jars, there is no need to flame sterilize between individual injection sites of the same jar.

The most commonly available way to flame sterilize would be an ordinary lighter, however, the butaine flame will leave a black residue on the needle. The black residue isn't really a big deal, but its ugly and can make it hard to tell when the needle is red hot.

The next best thing would be an alcohol lamp. I've tried to make my own acohol lamp many times out of canning jars and its just too much of a pain in the ass to get the flame just right. If the wick is too small, it'll take forever to heat the needle to red hot. If wick is too long, then it will pull the alcohol up out of the lamp through capillary action and start pooling on top of the jar, leading to a huge flaming fiasco lol.

My very favorite method of flame sterilizing my needles is a plumbers' propane torch. a mini pen torch will work too, but let's assume you don't have those handy at the moment... go with the lighter.

As RR says, "never contaminate a flame sterilized needle with alcohol"

Moving on...

Once you remove the foil, leave it off. Keeping the foil on the jars will suffocate them of GE, just as if they were in an air tight tupperware container. If you feel like it, you can put the foil back on while moving the jars to the place where they will be left to colonize. But once they are in their place, remove the foil.


--------------------
The Shaman

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