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InvisibleUna
controlleddemolition

Registered: 03/01/01
Posts: 970
Working with filter patch bags, some tips
    #928844 - 10/03/02 02:58 PM (21 years, 3 months ago)

Working with filter patch bags the easy way

Still many people think that it's impossible to sterilise sealed filter patch bags. They sterilise the substrate in an open bag and seal after spawning. This of course brings with it a great risk for introducing contaminants into the substrate.

We have had great succes with autoclaving sealed filter patch bags, however there are a few things you should know:

The bags expand during cooking which means that if the bottoms of the bags stand in water, they will push the water out of the cooker through the vent pipe. This can be prevented by placing the bags on top of a sturdy rack. The water level should be below the rack. This will mean that there is not much water in the cooker. Pressure has to be regulated precise so no excessive steam escape takes place.

Since the expanding bags may also block the vent pipe a rack of some sort should also be put on top of the bags to prevent this. If water comes out of the vent pipe this means that somehow there's too much water in the cooker and that the expanding bags can push the water out.

There should be enough bags in the autoclave. If the expanded volume of the bags is smaller than the volume of the cooker they will burst. Because they all expand during cooking they prevent each other from bursting.

As much air as possible is pushed out during sealing of the bags. This makes it easier to put them in the cooker and allows for better steam buildup.

Water and dry ingredients can be conveniently combined and sterilised for an appropiate time.

A good sealer must be used, poor seals burst during autoclaving.

Sterilsation time for bags is 1.5 - 2.5 hours depending on the single volume/weight of the bags.

After sterilisation the bags are removed and shaken. Shaking is not always possible because many bags will come out with little air in the bag. This doesn't matter since the bags will inflate during incubation and can then be easily shaken.

Since the bag is closed now, inoculation can only take place by liquid injection. For spawning purposes small amounts of substrate are sterilised in bottles and inoculated with the desired strain. Extra care is taken to ensure the purity of these master cultures.

When these master are completely colonised they can be used for inoculations. Sterile water is added to the master bottle and this is shaken by hand for 10 minutes. This broth is left to stand for 5 minutes after which the liquid can be poured back into the bottle that contained the water. This liquid is poured into a sterile container from which a 50 ml syringe can be filled.

This syringe is used to inoculate the bags. The injection site is cleaned with an alcohol soaked towel and then pierced with the needle. Liquid inoculant is injected into the substrate. The hole is shut with a drop of hot glue from a glue gun.

After the glue has hardened the bags should be shaken (again, if possible) and can then be incubated. Shaken can take place as soon as the bags have inflated (after some mycelial growth, often after a few days.)

Bags can be colonised in 10-15 days depending on many factors.


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InvisibleJoshua
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Re: Working with filter patch bags, some tips [Re: Una]
    #929789 - 10/03/02 09:29 PM (21 years, 3 months ago)

Thanks for the tips Una.

Joshua


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InvisibleOlgualion
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Re: Working with filter patch bags, some tips [Re: Una]
    #930715 - 10/04/02 01:17 AM (21 years, 3 months ago)

Yes! Thanks for the info!


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Offlineblackout
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Re: Working with filter patch bags, some tips [Re: Olgualion]
    #931222 - 10/04/02 04:52 AM (21 years, 3 months ago)

i make my own self healing patches by putting a bit of silicon on the filter patch and injecting through it. i used to put it on the plastic but found some of them fell off. it sticks much better to the filter patch. just make sure you dont cover it up totally!


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InvisibleUna
controlleddemolition

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Re: Working with filter patch bags, some tips [Re: blackout]
    #938438 - 10/07/02 01:19 AM (21 years, 3 months ago)

Quote:

i make my own self healing patches by putting a bit of silicon on the filter patch and injecting through it. i used to put it on the plastic but found some of them fell off. it sticks much better to the filter patch. just make sure you dont cover it up totally!




I like that idea, i'm going to try it :smile: 


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Offlinetplague
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Re: Working with filter patch bags, some tips [Re: Una]
    #1150715 - 12/18/02 04:10 AM (21 years, 1 month ago)

2 things -

1- i really like your filter patch bag post, maybe you should recomend it go into the FAQ ?

2 - there is something (i think) important that is a big general
"There should be enough bags in the autoclave."
I really have no idea how much grain, or how many bags to use, could you be more descriptive with this?

this hobby is so fun!


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OfflineRaadt
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Re: Working with filter patch bags, some tips [Re: tplague]
    #1150777 - 12/18/02 04:35 AM (21 years, 1 month ago)

you can post it in the FAQ contributions forum, at the bottom of the forum list.


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Raadt

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InvisibleUna
controlleddemolition

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Re: Working with filter patch bags, some tips [Re: tplague]
    #1151427 - 12/18/02 09:03 AM (21 years, 1 month ago)

There should be enough bags in the autoclave. If the expanded volume of the bags is smaller than the volume of the cooker they will burst. Because they all expand during cooking they prevent each other from bursting.


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OfflineBlueBruiser
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Re: Working with filter patch bags, some tips [Re: Una]
    #1159527 - 12/21/02 04:35 AM (21 years, 1 month ago)

Great post, I dont see many people using filter patch bags and I think they are much easier to use than jars. They hold way more substrate and are cheap and disposable. I also sterilize my bags sealed and it works just great. once again great post.


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OfflineSeussA
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Re: Working with filter patch bags, some tips [Re: Una]
    #1160301 - 12/21/02 12:25 PM (21 years, 1 month ago)

There should be enough bags in the autoclave. If the expanded volume of the bags is smaller than the volume of the cooker they will burst. Because they all expand during cooking they prevent each other from bursting.

I use a pelton and crane autoclave. The heating elements for the pressure chamber are along the sides. If a let a bag touch the sides at all, it will melt. I have to keep the bag in a bread pan and tie the pan up with cotton string to keep the bags from expanding into the sides and melting. After PCing, I remove the bag and let it cool in a flow hood... innoculate and impulse seal... no contamination.

I don't doubt that you have had success, Una... but my setup with the bags I am using (same style as from fungi perfect) I melt or burst every time if sealed.


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OfflineISH
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Re: Working with filter patch bags, some tips [Re: Seuss]
    #1227610 - 01/17/03 06:57 PM (21 years, 15 days ago)

Sounds like alot of work.


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OfflineFungusmaximusFM
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Re: Working with filter patch bags, some tips [Re: ISH]
    #1570336 - 05/22/03 08:13 PM (20 years, 8 months ago)

How do you do grain in these bags?
All I ever end up with is a bag of grain that has lots of excess water all over the grains they always end up in wet spot contaminations.


I have been PCing grain jars and filling the bags under the flowhood with the finished grain and re PCing the whole bag.
This is a PITA and sometimes it doesnt always work out.

Thanks.


FM


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Invisiblemicro
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Re: Working with filter patch bags, some tips [Re: FungusmaximusFM]
    #1570827 - 05/22/03 11:19 PM (20 years, 8 months ago)

The bags shouldn't be immersed in the water (just enough water so the p.c. definatly won't run out.) Also, I fold them down like 3 times and paperclip so they're a bit loose, but keep them in a conformation where the fold is on top.

The trick, IMO, is to fold them really neatly and space them in the p.c. so they will stay in the proper conformation (with as little air as possible;) this makes it so you can get a perfect seal when they come out. If they stay folded like three times (1/4") when you take them out, they should still be sterile for a little bit due to Pasteur's principle (nothing will really get in through all the folds, as long as you don't leave it like that.)

I enjoyed this post; I tried p.c.ing a sealed bag before, but it was only one bag in a tiny p.c. and it inflated and covered the regulator and the p.c. blew a gasket. The multiple bags and canning tray on the top are good ideas, though. It would make life a lot easier since I've had to be so careful with folding them to get a good seal.

Also (sorry if I already said this) -- you can inject a good amount of sterile water in a bag from the top (w/ alcohol and seal hole, of course) into a corner of the bag and mix a bit of the sterile colonized substrate with it and pull it back up with another sterile seringe. The only problem with this is you have to squirt the water in so that it only goes into a corner and doesn't touch the substrate, otherwise the substrate will absorb most of the water and you'll get little innoculum + wet mycelium. Also, no need to use that much col.'d substrate or the same problem will occur. If done right, though (easier with 2 people) you can make quick + easy sterile innoculum without the need of a flow-hood/glovebox, and then just take the rest of the col.'d substrate and case it, or whatever you will.

Thanks for the post!

--
Micro


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InvisiblePrisoner#1
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Re: Working with filter patch bags, some tips [Re: Una]
    #1575285 - 05/24/03 08:57 PM (20 years, 8 months ago)

Quote:

There should be enough bags in the autoclave. If the expanded volume of the bags is smaller than the volume of the cooker they will burst. Because they all expand during cooking they prevent each other from bursting.





I prevent them bursting by placing the second rack from my AA model on the top of the bags, place small jars or anything to take up space on top of that rack...it works pretty well....

P#1


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InvisiblePrisoner#1
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Re: Working with filter patch bags, some tips [Re: micro]
    #1575308 - 05/24/03 09:05 PM (20 years, 8 months ago)

The method I had used was soak the grain for around 7-12 hours, fold the top over just once and use box tape to secure it....or seal it and use the same methods

P#1


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OfflineBaby_Hitler
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Re: Working with filter patch bags, some tips [Re: Prisoner#1]
    #1575380 - 05/24/03 09:58 PM (20 years, 8 months ago)

I thought about just tieing them up real tight in a pillowcase.


--------------------
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Anonymous

Re: Working with filter patch bags, some tips [Re: Baby_Hitler]
    #1575774 - 05/25/03 01:32 AM (20 years, 8 months ago)

Once I forced some bags with all the air pushed out of them into a laundry bag, then put small weight over it to keep the bags flat, then sealed the bags shut.

The bags exploded inside.


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OfflineBaby_Hitler
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Re: Working with filter patch bags, some tips [Re: daba]
    #1575897 - 05/25/03 04:25 AM (20 years, 8 months ago)

How full was the pressure cooker? How tight was the bag? I was going to tie the bag tight so that there was zero room for them to expand.


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OfflineWorkmanV
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Re: Working with filter patch bags, some tips [Re: daba]
    #1576120 - 05/25/03 10:16 AM (20 years, 8 months ago)

There are new bags at MycoBag that can be sealed before sterilizing without bursting.


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Anonymous

Re: Working with filter patch bags, some tips [Re: Workman]
    #1576800 - 05/25/03 06:59 PM (20 years, 8 months ago)

Baby Hitler:
I didn't use a pressure cooker I used an AA25X at 21 PSI @ 260F.
At those temperatures, bags melt.

Workman:
The MycoBag Pre-sealable bags are outrageously high priced.


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OfflineWorkmanV
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Re: Working with filter patch bags, some tips [Re: daba]
    #1577171 - 05/25/03 09:40 PM (20 years, 8 months ago)

Yeah, they are high. I guess if they didn't have that selfhealing port they would be cheaper. Might ask if they have any without the port.


--------------------
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Anonymous

Re: Working with filter patch bags, some tips [Re: Workman]
    #1577213 - 05/25/03 09:53 PM (20 years, 8 months ago)

Indeed.
If you are ready to handle 1000's of bags, you can get them with free shipping and very low prices at www.unicornbags.com.
I believe that is where Mycobag gets their bags from, as well as SporeWorks.


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InvisibleUna
controlleddemolition

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Re: Working with filter patch bags, some tips [Re: Una] * 1
    #1577703 - 05/26/03 02:04 AM (20 years, 8 months ago)

I hope this image makes things clearer.



Note that the water level is BELOW the rack.


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InvisiblePrisoner#1
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Re: Working with filter patch bags, some tips [Re: Workman]
    #1577840 - 05/26/03 05:24 AM (20 years, 8 months ago)

Quote:

There are new bags at MycoBag that can be sealed before sterilizing without bursting.




I have done that several times with the previous bags...bent the second rack all to hell but the bags held together....


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InvisiblePrisoner#1
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Re: Working with filter patch bags, some tips [Re: Baby_Hitler]
    #1577846 - 05/26/03 05:34 AM (20 years, 8 months ago)

Quote:

How full was the pressure cooker? How tight was the bag? I was going to tie the bag tight so that there was zero room for them to expand.




in a cloth it'll work....I tried it with tape a time or two and had a small blowout and the other 2 bags ripped when I removed the tape....but dropping them into a sturdy pillow case and taping around that has worked quite well....lots of tape...dont use express or priority mail tape....it bonds together


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InvisiblePrisoner#1
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Re: Working with filter patch bags, some tips [Re: daba]
    #1577849 - 05/26/03 05:38 AM (20 years, 8 months ago)

Quote:

Indeed.
If you are ready to handle 1000's of bags, you can get them with free shipping and very low prices at www.unicornbags.com.
I believe that is where Mycobag gets their bags from, as well as SporeWorks.





nope...I tried...they refered me to spore works and now to mycobag....they are aparently the US distributor for unicorn....and believe me....I'm not on a shoe string budget....10,000 bag order didnt sway them...maybe under a contract....who knows.... I will try again or contract it to another company....there are 2 more that produce spawn bags that I'm aware of...just have never ordered from them....


Edited by Prisoner#1 (05/26/03 05:42 AM)


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OfflineBaby_Hitler
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Re: Working with filter patch bags, some tips [Re: Prisoner#1]
    #1577968 - 05/26/03 08:53 AM (20 years, 8 months ago)

http://www.mycobags.com and http://www.mycobag.com are two different companies. Mycobag is the sporeworks spinoff.

Quote:

but dropping them into a sturdy pillow case and taping around that has worked quite well....lots of tape...dont use express or priority mail tape....it bonds together





I was just going to tie a knot in it.

I'm planning on making a high temperature silicone injection site too. Pre-puncturing the injection site may help relieve some pressure.


--------------------
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InvisiblePrisoner#1
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Re: Working with filter patch bags, some tips [Re: Baby_Hitler]
    #1579820 - 05/26/03 09:55 PM (20 years, 8 months ago)

3 out of 3 times I had coontacted unicorn bags I was given a referal to spore works, and the last was to mycobag.....they also provide to a few other but how that chain is linked is beyond me....but I started about a year and a half ago calling in reference to the products at unicorn....maybe they just dont wish to sell to me.



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InvisibleUna
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Re: Working with filter patch bags, some tips [Re: Prisoner#1]
    #1597799 - 06/01/03 06:58 AM (20 years, 7 months ago)

Here's a tip for people in Europe. "De Spore" is the european distributor of the Unicorn bags. This is the company from Peter Oei (author of the book "mushroom cultivation").

The cost (if i remember correctly) is 22 euro cents per bag.

Their homepage can be found HERE

English page about the spawnbags HERE


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Invisiblemicro
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Re: Working with filter patch bags, some tips [Re: Una]
    #1606308 - 06/03/03 09:22 PM (20 years, 7 months ago)

I just had an idea about making spore prints; never tried it, though. Why not just fruit 1 bag in vitro and take a cap or 3 or 4 off (in vitro, using the outside of the bag,) and flip the bag on its side, take the print on the bag and then fold that part and heat seal it? This should be completely sterile, and you could always just inject a little sterile DI water into your spore pouch, mix + pull up to make a seringe.

Just an idea....

--
Micro


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Re: Working with filter patch bags, some tips [Re: micro]
    #1607360 - 06/04/03 08:12 AM (20 years, 7 months ago)

It would be difficult to manipulate the caps through the bag, but you might could just turn it on it's side, and let the spores fall on the side of the bag. You wouldn't get a "print" exactly, but you would get plenty o spores.


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Invisiblesci33
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Re: Working with filter patch bags, some tips [Re: Baby_Hitler]
    #2567506 - 04/16/04 11:25 AM (19 years, 9 months ago)

Not to bring up an old thread but this is informative and I was wondering, couldn't you just take the sealed bag and pasteurize it? For grain/superspawn innoculations.


--------------------
Looking for:
Ps. weilii, Ps. samuiensis, Ps. hispanica, Pan. cyanescens (any strain I don't have)

Have:
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Offlinedebianlinux
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Re: Working with filter patch bags, some tips [Re: sci33]
    #2567634 - 04/16/04 12:10 PM (19 years, 9 months ago)

i'm confused, i think...

grains have to be sterilised, pasteurisation doesn't cut it with grains

i assume your talking about pasteurising straw in the bags to spawn and grow like a straw log?


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Invisiblesci33
AmbitiousScientist
Registered: 11/11/03
Posts: 186
Re: Working with filter patch bags, some tips [Re: debianlinux]
    #2567640 - 04/16/04 12:14 PM (19 years, 9 months ago)

Yes, I mean mixing up the dung/straw mixture in a filter patch bag, pasteurizing it in a water bath of 170 degrees for an hour or so, allowing it to cool down in a flowhood, then innoculating it with colonized grain or superspawn innoculation - and then sealing the bag.

That just seems like the quickest method to colonize the substrate to me rather than a syringe innoculation on sterilized substrate.


--------------------
Looking for:
Ps. weilii, Ps. samuiensis, Ps. hispanica, Pan. cyanescens (any strain I don't have)

Have:
Ps. cubensis, Ps. tampanensis, Pan. cyanescens (Australian, Tai-KS, Hawaiian, Jamaican, Ban Lipa Noi)

PM for a trade.


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