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Offlineeve.kalika

Registered: 09/11/14
Posts: 26
Last seen: 9 years, 1 month
Autoclave Times
    #20810993 - 11/08/14 06:37 PM (9 years, 2 months ago)

In regards to sterilizing rye grain jars: Every set of instructions I've found involving autoclaves has said to put the jars in the autoclave for 45 minutes. I've used an autoclave hundreds of times, and I've only ever put anything in for 15-20 minutes (then 20+ minutes to cool/de-pressurize). I've done agars, water, other fluids, empty containers, instruments, and pretty much anything you can imagine. The time is always 15-20 min. Is there a particular reason for all instructions on this site saying 45 min, or are people simply adapting instructions from PC teks and not taking into account the higher temps and pressure?

I don't see why mushroom substrate would need any longer than anything else, but I want to make sure I'm not missing something. I've spent the last hour searching this site, but I'm sorry if this was answered somewhere already.


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OfflinetaGyo
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Re: Autoclave Times [Re: eve.kalika]
    #20811011 - 11/08/14 06:42 PM (9 years, 2 months ago)

Is an autoclave different from a pressure cooker?
Does it have a PSI meter?


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Edited by taGyo (11/08/14 06:42 PM)


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Offlineinvitro

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Re: Autoclave Times [Re: eve.kalika]
    #20811135 - 11/08/14 07:16 PM (9 years, 2 months ago)

Mold and bacteria are harder to kill when they are on grains, the only explanation I've heard is that somehow the cleavage on the grain seed protects them from the heat.  You might want to wait for a better answer but experience has shown me that short sterilization times do not sterilize grains completely.


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OfflinetaGyo
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Re: Autoclave Times [Re: invitro]
    #20811148 - 11/08/14 07:19 PM (9 years, 2 months ago)

Quote:

invitro said:
Mold and bacteria are harder to kill when they are on grains, the only explanation I've heard is that somehow the cleavage on the grain seed protects them from the heat.  You might want to wait for a better answer but experience has shown me that short sterilization times do not sterilize grains completely.



Happy 1,000th post Invitro :patriot:
We had some scuffle in the past but good vibes and keep on keepin' on
:hippie:


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InvisibleMrGiraffe

Registered: 04/04/14
Posts: 3,149
Re: Autoclave Times [Re: taGyo]
    #20811365 - 11/08/14 08:00 PM (9 years, 2 months ago)

Everything you listed that you have put in an autoclave for 15-20 minutes doesn't have a lot of mass.  If you don't get the very center of the jar up to temp then it won't sterilize.


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Invisiblelipa

Registered: 07/24/07
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Re: Autoclave Times [Re: taGyo]
    #20811377 - 11/08/14 08:02 PM (9 years, 2 months ago)

Quote:

taGyo said:
Is an autoclave different from a pressure cooker?
Does it have a PSI meter?




A Sterilizer does not release steam and forms a vacuum upon cooldown.

A Pressure cooker releases steam during operation and sucks in air upon cooldown.


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OfflinetaGyo
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Re: Autoclave Times [Re: lipa]
    #20811392 - 11/08/14 08:04 PM (9 years, 2 months ago)

Quote:

lipa said:
Quote:

taGyo said:
Is an autoclave different from a pressure cooker?
Does it have a PSI meter?




A Sterilizer does not release steam and forms a vacuum upon cooldown.

A Pressure cooker releases steam during operation and sucks in air upon cooldown.



:mygoditsfullofstars:

Raised more questions. Is a sterilizer an autoclave? Invitro already explained to me the difference but what are you talking about?


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Offlineeve.kalika

Registered: 09/11/14
Posts: 26
Last seen: 9 years, 1 month
Re: Autoclave Times [Re: taGyo]
    #20811618 - 11/08/14 08:54 PM (9 years, 2 months ago)

Quote:

taGyo said:
Is an autoclave different from a pressure cooker?
Does it have a PSI meter?



Long story short: An autoclave is a pressure cooker on steroids. It does have a PSI metre. The pressure is higher and it gets to that pressure almost instantly.

Not all autoclaves are the same. Mine's pretty old school and it has a steam valve on the top like a pressure cooker, which I close as soon as the chamber fills with steam and then open only after it's cooled down, usually right before I open it (because A: I don't want a face full of steam, and B: I physically can't open it otherwise because it's vacuumed shut).

Quote:

MrGiraffe said:
Everything you listed that you have put in an autoclave for 15-20 minutes doesn't have a lot of mass.  If you don't get the very center of the jar up to temp then it won't sterilize.




Some of the things I autoclave are denser than a jar of grain. I'm too paranoid to be specific, but when I say instruments, I don't mean surgical tools. I mean things that are a lot larger and denser than a grain jar and do need to be sterilized to the centre, not just on the surface.

Although... grain jars are prepared in such a way that moisture content doesn't change inside and that interferes with steam penetration. I package things very loosely for this reason, but of course they get pretty wet. Anything I autoclave where moisture content is relevant is aqueous, which would be more easily affected by the pressure. Okay, so grain jars must need longer times because they're packaged and (relatively) dry.

Alright, I answered my own question... lol. Just had to think about it, I guess. Well, now it's here for anyone else with the same question.


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InvisiblePastywhyteMDiscord
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Re: Autoclave Times [Re: eve.kalika]
    #20811648 - 11/08/14 09:06 PM (9 years, 2 months ago)

I love my sterilizer and I still do just as long a run with grains as I did with a PC even tho I sterilize at a higher pressure. Some industrial autoclaves are different than sterilizers and are programmed to dump the pressure all at once. This is a problem as it will crack the jars, and pull the moisture out of the grains. Not sure what the OP is using tho.


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OfflineSiliconGhost
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Re: Autoclave Times [Re: Pastywhyte]
    #20811696 - 11/08/14 09:22 PM (9 years, 2 months ago)

FINALLY! A thread where I can give expert advice.

I use autoclaves exclusively for my grows. Check out my grow thread and you'll see one of my 'claves. I use the 45 minute gravity cycle for my jars, bags, etc and it has worked beautifully. Make sure it isn't a 45 min gravity cycle with a drying feature at the end. Even if that is the case, you don't need to worry too much.

Here is what I do:
  • 1. prep your jars or bags like normal.
  • 2. Grab an autoclave safe plastic tub and fill with 1.5 - 2 inches of water.
  • 3. place the tub with the jars in the autoclave and run your 45 min GRAVITY cycle
  • 4. There seems to have been some folks on here saying an autoclave will dry you out that is not the case.
  • 5. Allow the jars to cool before inoculating.
  • 6. I also use an autoclave liquid cycle to sterilize all of my agar as well.
  • 7. Autoclave's are used for fungi spawn in labs all over the world, but not available to most people. Hence the reason there isn't a lot of info available here.


Good luck man. PM me if you have any more questions. I posted the link above, but here is a link for a recent grow I did with stuff all through an autoclave.
http://www.shroomery.org/forums/showflat.php/Number/20595126


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OfflineSiliconGhost
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Re: Autoclave Times [Re: taGyo]
    #20811730 - 11/08/14 09:31 PM (9 years, 2 months ago)

Quote:

taGyo said:
Quote:

lipa said:
Quote:

taGyo said:
Is an autoclave different from a pressure cooker?
Does it have a PSI meter?




A Sterilizer does not release steam and forms a vacuum upon cooldown.

A Pressure cooker releases steam during operation and sucks in air upon cooldown.



:mygoditsfullofstars:

Raised more questions. Is a sterilizer an autoclave? Invitro already explained to me the difference but what are you talking about?




There is a lot of confusion on here about sterilizers vs autoclaves.

Not all sterilizers are autoclaves, but all autoclaves function as sterilizers. Old sterilizers were kindof like an autoclave, but there is a functional difference. In a lab setting, there are sterilizers and autoclave. Sterilizers are generally accepted to be "dry-heat sterilizers" and do not use steam to sterilize their contents.

Autoclaves are generally accepted to be large appliances that are sometimes counter-top but are generally a large wall/floor unit used to sterilize large amounts of things. They remove all of the air from the chamber and increase pressure of the chamber to sterilize the media. They have multiple cycles for different media etc.

There are stove-top autoclaves which are also known as steam sterilizers. I think this is also what causes a lot of confusion on here.


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OfflineKizzle
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Re: Autoclave Times [Re: eve.kalika]
    #20811845 - 11/08/14 10:01 PM (9 years, 2 months ago)

Quote:

eve.kalika said:
In regards to sterilizing rye grain jars: Every set of instructions I've found involving autoclaves has said to put the jars in the autoclave for 45 minutes. I've used an autoclave hundreds of times, and I've only ever put anything in for 15-20 minutes (then 20+ minutes to cool/de-pressurize). I've done agars, water, other fluids, empty containers, instruments, and pretty much anything you can imagine. The time is always 15-20 min. Is there a particular reason for all instructions on this site saying 45 min, or are people simply adapting instructions from PC teks and not taking into account the higher temps and pressure?

I don't see why mushroom substrate would need any longer than anything else, but I want to make sure I'm not missing something. I've spent the last hour searching this site, but I'm sorry if this was answered somewhere already.



The reason is heat penetration.  Substrate can't have the stream heat it directly or it ruins the moisture content. A substrate jar/bag  contains a lot of air which acts as an insulator since the steam doesn't flows through it.


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OfflineSiliconGhost
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Re: Autoclave Times [Re: Kizzle]
    #20811959 - 11/08/14 10:40 PM (9 years, 2 months ago)

Quote:

Kizzle said:
Quote:

eve.kalika said:
In regards to sterilizing rye grain jars: Every set of instructions I've found involving autoclaves has said to put the jars in the autoclave for 45 minutes. I've used an autoclave hundreds of times, and I've only ever put anything in for 15-20 minutes (then 20+ minutes to cool/de-pressurize). I've done agars, water, other fluids, empty containers, instruments, and pretty much anything you can imagine. The time is always 15-20 min. Is there a particular reason for all instructions on this site saying 45 min, or are people simply adapting instructions from PC teks and not taking into account the higher temps and pressure?

I don't see why mushroom substrate would need any longer than anything else, but I want to make sure I'm not missing something. I've spent the last hour searching this site, but I'm sorry if this was answered somewhere already.



The reason is heat penetration.  Substrate can't have the stream heat it directly or it ruins the moisture content. A substrate jar/bag  contains a lot of air which acts as an insulator since the steam doesn't flows through it.




^^ :whathesaid:

This guy is spot on. With spawn in jars it's not an issue. Only with substrates it can be an issue. I do all of my jars on 45 minute cycles to eliminate any threat of endospores. The longer cycle ensures that your spawn is completely sterile.

In theory you could "pasteurize" spawn in an autoclave short cycle, but I would advise strongly against it. The only substrate I would ever consider throwing in an autoclave would be straight CVG and I mean STRAIGHT. Coir is pretty contam resistant, but if you get far outside just CVG you're asking for trouble if you accidentally sterilize the substrate.

Spawn bags are fine in an autoclave, but only the ones made out of Polypropylene. MAKE VERY DAMN SURE that you're on a setting at 121c and not 134c. 134c will fuck certain plastics, and I mean fuck them hard.


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Offlineeve.kalika

Registered: 09/11/14
Posts: 26
Last seen: 9 years, 1 month
Re: Autoclave Times [Re: SiliconGhost]
    #20812025 - 11/08/14 11:06 PM (9 years, 2 months ago)

Quote:

The reason is heat penetration.  Substrate can't have the stream heat it directly or it ruins the moisture content. A substrate jar/bag  contains a lot of air which acts as an insulator since the steam doesn't flows through it.



I did actually realize that, if you read my second post (not the original one). Thanks for confirming. :smile:

Quote:

SiliconGhost said:
FINALLY! A thread where I can give expert advice.

I use autoclaves exclusively for my grows. Check out my grow thread and you'll see one of my 'claves. I use the 45 minute gravity cycle for my jars, bags, etc and it has worked beautifully. Make sure it isn't a 45 min gravity cycle with a drying feature at the end. Even if that is the case, you don't need to worry too much.

Here is what I do:
  • 1. prep your jars or bags like normal.
  • 2. Grab an autoclave safe plastic tub and fill with 1.5 - 2 inches of water.
  • 3. place the tub with the jars in the autoclave and run your 45 min GRAVITY cycle
  • 4. There seems to have been some folks on here saying an autoclave will dry you out that is not the case.
  • 5. Allow the jars to cool before inoculating.
  • 6. I also use an autoclave liquid cycle to sterilize all of my agar as well.
  • 7. Autoclave's are used for fungi spawn in labs all over the world, but not available to most people. Hence the reason there isn't a lot of info available here.


Good luck man. PM me if you have any more questions. I posted the link above, but here is a link for a recent grow I did with stuff all through an autoclave.
http://www.shroomery.org/forums/showflat.php/Number/20595126



Thanks! So the water bath promotes heat penetration?
This autoclave is not at all automated. Very unsophisticated :p. There are no cycles and there is no drying feature. There's a steam valve coming directly from a boiler, so I control temp/pressure simply by adjusting that valve. The pressure release valve is also manual, so I can control all of that and do a "gravity cycle" by exhausting faster.

Quote:

Spawn bags are fine in an autoclave, but only the ones made out of Polypropylene. MAKE VERY DAMN SURE that you're on a setting at 121c and not 134c. 134c will fuck certain plastics, and I mean fuck them hard.



I better stick with jars. I don't have settings, I just adjust the valve and watch the dial. Not very exact.


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OfflineSiliconGhost
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Re: Autoclave Times [Re: eve.kalika]
    #20812101 - 11/08/14 11:31 PM (9 years, 2 months ago)

Quote:

eve.kalika said:
Quote:

The reason is heat penetration.  Substrate can't have the stream heat it directly or it ruins the moisture content. A substrate jar/bag  contains a lot of air which acts as an insulator since the steam doesn't flows through it.



I did actually realize that, if you read my second post (not the original one). Thanks for confirming. :smile:

Quote:

SiliconGhost said:
FINALLY! A thread where I can give expert advice.

I use autoclaves exclusively for my grows. Check out my grow thread and you'll see one of my 'claves. I use the 45 minute gravity cycle for my jars, bags, etc and it has worked beautifully. Make sure it isn't a 45 min gravity cycle with a drying feature at the end. Even if that is the case, you don't need to worry too much.

Here is what I do:
  • 1. prep your jars or bags like normal.
  • 2. Grab an autoclave safe plastic tub and fill with 1.5 - 2 inches of water.
  • 3. place the tub with the jars in the autoclave and run your 45 min GRAVITY cycle
  • 4. There seems to have been some folks on here saying an autoclave will dry you out that is not the case.
  • 5. Allow the jars to cool before inoculating.
  • 6. I also use an autoclave liquid cycle to sterilize all of my agar as well.
  • 7. Autoclave's are used for fungi spawn in labs all over the world, but not available to most people. Hence the reason there isn't a lot of info available here.


Good luck man. PM me if you have any more questions. I posted the link above, but here is a link for a recent grow I did with stuff all through an autoclave.
http://www.shroomery.org/forums/showflat.php/Number/20595126



Thanks! So the water bath promotes heat penetration?
This autoclave is not at all automated. Very unsophisticated :p. There are no cycles and there is no drying feature. There's a steam valve coming directly from a boiler, so I control temp/pressure simply by adjusting that valve. The pressure release valve is also manual, so I can control all of that and do a "gravity cycle" by exhausting faster.

Quote:

Spawn bags are fine in an autoclave, but only the ones made out of Polypropylene. MAKE VERY DAMN SURE that you're on a setting at 121c and not 134c. 134c will fuck certain plastics, and I mean fuck them hard.



I better stick with jars. I don't have settings, I just adjust the valve and watch the dial. Not very exact.





H'okay I got this. However you control it you want to do the following:

  • 1. Internal chamber pressure of 15 psi
  • 2. Internal temp of AT least 121˚C /250˚F
  • 3. Hold this pressure and temp for 30-60 min, shoot for 45min.
  • 4. The difference between gravity cycles and liquid cycles is how the chamber exhaust is handled. Liquid cycles slowly vent the chamber so as not to boil the liquids (slow-exhaust). Gravity cycles (for solids or glassware etc) release the exhaust valve and the chamber exhaust is dumped quickly (fast-exhaust). Just like you said.


Here is a good link that explains things as I have done. You'll be fine OP. You've got the understanding of how an autoclave and that's all that is required to make it work for yah. There's just a big lack of info on this site simply because most folks don't have access to a legit autoclave outside of lab or commercial settings.

The water bath isn't as much for penetration as it is for even heating. Also helps when the glass jars expand and contract with the heat. Just like when you autoclave glassware and don't put them directly on the floor of the autoclave. Yah dig?

Check out my thread for pics.


--------------------
---------SILICONGHOST---------

All posts involve edible mushroom varieties. No controlled substances

Magnetic Stirring Hot Plates for Sale/Trade. :pm: Me


Edited by SiliconGhost (11/08/14 11:35 PM)


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Offlineeve.kalika

Registered: 09/11/14
Posts: 26
Last seen: 9 years, 1 month
Re: Autoclave Times [Re: SiliconGhost]
    #20813081 - 11/09/14 08:41 AM (9 years, 2 months ago)

Thanks for the info! Super helpful. :grin:


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OfflineKizzle
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Re: Autoclave Times [Re: eve.kalika]
    #20813534 - 11/09/14 10:48 AM (9 years, 2 months ago)

45 minutes is really the bare minimum for spawn and your success can vary depending on other factors. I actually don't of know of anyone that uses less than 60 minutes at 15 psi. I would familiarize yourself well with all the signs of bacteria on grain because you may need to increase it to 60 or even longer, especially with larger jars/bags.

PC times and autoclave times are actually the same not counting the extra time a PC takes to reach full pressure.


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Offlineeve.kalika

Registered: 09/11/14
Posts: 26
Last seen: 9 years, 1 month
Re: Autoclave Times [Re: Kizzle]
    #20813863 - 11/09/14 12:09 PM (9 years, 2 months ago)

This autoautoclave actually operates at closer to 30 psi, so a shorter time can be used.

Ultimately, there's only one way to find out. I can leave a blank control to ensure it's not contaminated from inadequate sterilizing.

How long after sterilizing can I keep a grain jar?


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OfflineSiliconGhost
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Re: Autoclave Times [Re: eve.kalika]
    #20814104 - 11/09/14 12:59 PM (9 years, 2 months ago)

Quote:

eve.kalika said:
This autoautoclave actually operates at closer to 30 psi, so a shorter time can be used.

Ultimately, there's only one way to find out. I can leave a blank control to ensure it's not contaminated from inadequate sterilizing.

How long after sterilizing can I keep a grain jar?




15 is the minimum. I run 45 minutes cycles at 30 PSI and 121˚C. I've not had one contam on it's on after autoclaved. My contams are usually the result of me cracking the lids to inoculate. I now work in front of a flow hood in one of the other in vitro labs down the hall.


--------------------
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Offlineeve.kalika

Registered: 09/11/14
Posts: 26
Last seen: 9 years, 1 month
Re: Autoclave Times [Re: SiliconGhost]
    #20814418 - 11/09/14 01:59 PM (9 years, 2 months ago)

I've got a flow hood, too. I'm wondering how long a jar will last.


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Edited by eve.kalika (12/07/14 08:04 PM)


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