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trentallica
Lord Trentnar


Registered: 05/01/06
Posts: 797
Loc: virgina
Last seen: 8 years, 8 months
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will an autoclave destroy nutrients in a growth medium???
#5658601 - 05/21/06 11:14 PM (17 years, 8 months ago) |
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i don't have an pressure cooked, but i do have an autoclave and have been using it. an autoclave dose not have a max pressure or temp setting. it just gets as hot as it going to get, and gets as much pressure as its going to get. it is used to sterilize med equipment so there in no need for max limits. when i use it pressure gets as high as 30 psi, and temp over 260 F. everything i have inoculated past a certain point shown no sign of growth. ok to be fair some of it was inoculated today, but still. I'm trying to think what i did differently the first two times that I'm not doing now. one thing that comes to mind is the autoclave. my first two attempts both came from the same autoclave cycle. the cycles that i have used after that have all been on longer cycles just to "be on the safe side", so they have all had longer exposure to ever higher temps , and higher psi then the first cycle. i know nutrients are not alive, but i know proteins ans amino acids are involved. could the autoclave be converting the nutrients to an unusable state??????
-------------------- The only ones for me are the mad ones..........
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FooMan



Registered: 02/02/05
Posts: 8,957
Loc: Earth
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Re: will an autoclave destroy nutrients in a growth medium??? [Re: trentallica]
#5658640 - 05/21/06 11:23 PM (17 years, 8 months ago) |
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Quote:
trentallica said: i don't have an pressure cooked, but i do have an autoclave and have been using it. an autoclave dose not have a max pressure or temp setting. it just gets as hot as it going to get, and gets as much pressure as its going to get. it is used to sterilize med equipment so there in no need for max limits. when i use it pressure gets as high as 30 psi, and temp over 260 F. everything i have inoculated past a certain point shown no sign of growth. ok to be fair some of it was inoculated today, but still. I'm trying to think what i did differently the first two times that I'm not doing now. one thing that comes to mind is the autoclave. my first two attempts both came from the same autoclave cycle. the cycles that i have used after that have all been on longer cycles just to "be on the safe side", so they have all had longer exposure to ever higher temps , and higher psi then the first cycle. i know nutrients are not alive, but i know proteins ans amino acids are involved. could the autoclave be converting the nutrients to an unusable state??????
I'm not sure about the nutrient loss, but at 30psi you would cut the sterilization time in half, not go longer. It could be many things that are causing you to fail: bad spores, inoculating while the jars are still hot, releasing pressure manually instead of letting it drop on it's own, improper moisture content, high/low incubation temps, improper substrate mix, blah, blah, blah.
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Quick WBS Prep
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trentallica
Lord Trentnar


Registered: 05/01/06
Posts: 797
Loc: virgina
Last seen: 8 years, 8 months
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Re: will an autoclave destroy nutrients in a growth medium??? [Re: FooMan]
#5658706 - 05/21/06 11:39 PM (17 years, 8 months ago) |
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hmmm spores. that would be the other thing the jars that are working have in common that are all the same spore. i cant use logic to rule out inoculating too soon, or pressure release was differnt, but the working ones and the non working ones all have the same environment.
i still need to know about the autoclave though???
-------------------- The only ones for me are the mad ones..........
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BooNanZa
Microbiologist
Registered: 07/05/05
Posts: 16
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Re: will an autoclave destroy nutrients in a growth medium??? [Re: trentallica]
#5658792 - 05/22/06 12:00 AM (17 years, 8 months ago) |
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I too was wondering this. I have access to an autoclave at work, and it has two settings, LIQUID (for liquid cultures and garbage bags) and GRAVITY (for beakers, etc.) and wanted to know if I could throw some spawn bags in there since they won't fit my PC. It also goes to about 30psi, with a run of 35 minutes (20 minutes sterilize, 15 minutes dry)
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trentallica
Lord Trentnar


Registered: 05/01/06
Posts: 797
Loc: virgina
Last seen: 8 years, 8 months
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Re: will an autoclave destroy nutrients in a growth medium??? [Re: BooNanZa]
#5659439 - 05/22/06 08:49 AM (17 years, 8 months ago) |
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im looking more into it , and doing a little experamentation as well. ill post to this topic again when i know more.
-------------------- The only ones for me are the mad ones..........
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Edgekrusher
God
Registered: 10/10/05
Posts: 674
Last seen: 17 years, 1 month
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Re: will an autoclave destroy nutrients in a growth medium??? [Re: trentallica]
#5659518 - 05/22/06 09:25 AM (17 years, 8 months ago) |
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If you pressure cook for an hour at 15psi. You'd only need 30mins at 30psi.
I wouldn't imagine its the autoclave messing things up. Shit, if I had one I'd retire my PC right this moment. Give the innoculations atleat 5 days incubating if using MS innoculations. If not growth after 5-7 days, then worry.
Patience is a virtue needed for mycology.
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Tweexican
Clit Commander

Registered: 11/06/03
Posts: 657
Last seen: 5 years, 6 months
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Re: will an autoclave destroy nutrients in a growth medium??? [Re: BooNanZa]
#5659775 - 05/22/06 11:25 AM (17 years, 8 months ago) |
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Quote:
BooNanZa said: I too was wondering this. I have access to an autoclave at work, and it has two settings, LIQUID (for liquid cultures and garbage bags) and GRAVITY (for beakers, etc.) and wanted to know if I could throw some spawn bags in there since they won't fit my PC. It also goes to about 30psi, with a run of 35 minutes (20 minutes sterilize, 15 minutes dry)
where do you work???!!!
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