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InvisibleUna
controlleddemolition

Registered: 03/01/01
Posts: 970
Working with filter patch bags, some tips
    #928844 - 10/03/02 04:58 PM (18 years, 10 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 11:29 PM (18 years, 10 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 03:17 AM (18 years, 10 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 06:52 AM (18 years, 9 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

Registered: 03/01/01
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Re: Working with filter patch bags, some tips [Re: blackout]
    #938438 - 10/07/02 03:19 AM (18 years, 9 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 06:10 AM (18 years, 7 months 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 06:35 AM (18 years, 7 months ago)

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


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InvisibleUna
controlleddemolition

Registered: 03/01/01
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Re: Working with filter patch bags, some tips [Re: tplague]
    #1151427 - 12/18/02 11:03 AM (18 years, 7 months 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 06:35 AM (18 years, 7 months 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 02:25 PM (18 years, 7 months 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 08:57 PM (18 years, 6 months 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 10:13 PM (18 years, 2 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 - 05/23/03 01:19 AM (18 years, 2 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 10:57 PM (18 years, 2 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 11:05 PM (18 years, 2 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 11:58 PM (18 years, 2 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 03:32 AM (18 years, 2 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 06:25 AM (18 years, 2 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 12:16 PM (18 years, 2 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 08:59 PM (18 years, 2 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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