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Doc_T
Random Dude




Registered: 03/06/09
Posts: 42,395
Loc: Colorado
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Re: Losing the battle with bacrteria [Re: fngbronco]
#14019445 - 02/24/11 11:28 AM (13 years, 9 days ago) |
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Quote:
fngbronco said: How should I inoculate ones with polyfil (no money for a while) and how tight should it be placed in the holes?
You can shoot past the polyfil puff if you're careful. Or make silicone injection ports. I fold a little puff over, then reach through with piers and pull it. Too tight to stuff by hand.
-------------------- You make it all possible. Doesn't it feel good?
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fngbronco
Monkey Man



Registered: 09/26/10
Posts: 2,877
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Re: Losing the battle with bacrteria [Re: Doc_T]
#14019497 - 02/24/11 11:40 AM (13 years, 9 days ago) |
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K sweet! I'll be starting this whole deal over tomorrow! Gonna empty out and wash and pc all my jars so they're all sterile and clean today! Buying new metal lids and rings and drilling those as well. Stupid ass bacteria out smarting me....
-------------------- I challenge you to challenge yourself more! When you feel complacent and ready to hang it up, challenge yourself to get over it! If you fail, don't look at it as you didn't succeed, look at it as you would a rock face you're trying to climb. Stand back, wayyyy back, and look at it and plot another path. If you can't find one, shuffle down the way a little, a little change of scenery or a view from a different angle may give you the insight you need. Anything I state is relayed information from a friend of a friend and should be viewed as completely fictitious. I do not partake in any illegal or grey-area-of-the-law activities, but do have lots of friends who may or may not. -fngbronco Pill Divider Agar Tek
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Doc_T
Random Dude




Registered: 03/06/09
Posts: 42,395
Loc: Colorado
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Re: Losing the battle with bacrteria [Re: fngbronco]
#14019512 - 02/24/11 11:42 AM (13 years, 9 days ago) |
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Just don't keep shooting with the same dirty syringe. Get some agar while you're out.
-------------------- You make it all possible. Doesn't it feel good?
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fngbronco
Monkey Man



Registered: 09/26/10
Posts: 2,877
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Re: Losing the battle with bacrteria [Re: Doc_T]
#14019959 - 02/24/11 01:20 PM (13 years, 9 days ago) |
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I have agar but it grows bacteria as well. The syringes are clean and the myc is strong (took over some trich on one plate). I've had 3 successful agar plates and even the closed sealed ones will contam (possibly from condensation). Once my glove box has the kinks worked out, to where's I can leave an open plate in there for a couple uses and not get contams, then I'll start at it again. I've just washed all my glass in NABC and am pressure sterilizing at 10psi for 15 min. I'm wondering if my bare fingers on the tape I put over the holes may have attributed as well. I've shot up pf jars with the same solutions with no contams.
-------------------- I challenge you to challenge yourself more! When you feel complacent and ready to hang it up, challenge yourself to get over it! If you fail, don't look at it as you didn't succeed, look at it as you would a rock face you're trying to climb. Stand back, wayyyy back, and look at it and plot another path. If you can't find one, shuffle down the way a little, a little change of scenery or a view from a different angle may give you the insight you need. Anything I state is relayed information from a friend of a friend and should be viewed as completely fictitious. I do not partake in any illegal or grey-area-of-the-law activities, but do have lots of friends who may or may not. -fngbronco Pill Divider Agar Tek
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Doc_T
Random Dude




Registered: 03/06/09
Posts: 42,395
Loc: Colorado
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Re: Losing the battle with bacrteria [Re: fngbronco]
#14022874 - 02/24/11 10:21 PM (13 years, 8 days ago) |
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Quote:
fngbronco said: I've had 3 successful agar plates and even the closed sealed ones will contam (possibly from condensation).
No. Contams come from contams. Contams do not come from sterile water. Your pressure cooking procedure is faulty. Review correct technique and practice doing it right.
You should be able to leave an agar plate alone and have it stay sterile. End of story.
-------------------- You make it all possible. Doesn't it feel good?
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fngbronco
Monkey Man



Registered: 09/26/10
Posts: 2,877
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Re: Losing the battle with bacrteria [Re: Doc_T]
#14022908 - 02/24/11 10:28 PM (13 years, 8 days ago) |
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I'm sayin the contams come from the glove box. My sterility was lacking there. All the jars are have been cleaned thoroughly and tomorrow will be my glove box nuking day. Gonna make some agar this weekend and pour test plates. What's the best way to clean things before putting them in the glove box, so as to not drag in contams?
-------------------- I challenge you to challenge yourself more! When you feel complacent and ready to hang it up, challenge yourself to get over it! If you fail, don't look at it as you didn't succeed, look at it as you would a rock face you're trying to climb. Stand back, wayyyy back, and look at it and plot another path. If you can't find one, shuffle down the way a little, a little change of scenery or a view from a different angle may give you the insight you need. Anything I state is relayed information from a friend of a friend and should be viewed as completely fictitious. I do not partake in any illegal or grey-area-of-the-law activities, but do have lots of friends who may or may not. -fngbronco Pill Divider Agar Tek
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biologys
Mycologist in Trainning




Registered: 12/21/09
Posts: 4,622
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Re: Losing the battle with bacrteria [Re: fngbronco]
#14022915 - 02/24/11 10:29 PM (13 years, 8 days ago) |
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just a simple bleach/water mixture, then soap/water is all I do.
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Doc_T
Random Dude




Registered: 03/06/09
Posts: 42,395
Loc: Colorado
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Re: Losing the battle with bacrteria [Re: fngbronco]
#14022931 - 02/24/11 10:33 PM (13 years, 8 days ago) |
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You are doing something wrong. Is it a glovebox or still air box? Either way, wash it out with soap and water. Put items in, close it, wait 15 minutes.
Now, wash hands and glove up. Then go to work.
Leave one dish untouched. If it contams, bad PCing. Open one dish in the glovebox, but don't inoculate it. Open one dish, drag you flamed scalpel through it, close it up
It's gotta be you touching something somewhere. Wish you could shoot a movie, we could tell you in a second.
-------------------- You make it all possible. Doesn't it feel good?
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Doc_T
Random Dude




Registered: 03/06/09
Posts: 42,395
Loc: Colorado
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Re: Losing the battle with bacrteria [Re: Doc_T]
#14022941 - 02/24/11 10:34 PM (13 years, 8 days ago) |
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Wait, wait, wait. You said your syringes are clean? You've never grown a clean culture, how do you know?
-------------------- You make it all possible. Doesn't it feel good?
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Mikeallojee
Coolaid smile



Registered: 10/02/09
Posts: 897
Loc: SW WA
Last seen: 11 years, 19 days
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Re: Losing the battle with bacrteria [Re: Doc_T]
#14023313 - 02/24/11 11:49 PM (13 years, 8 days ago) |
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Plastic lids can be difficult but they can also be handy if used correctly. I have begun to flip the lids on the jars. When used upside down you cant beat them. They slide around a bit but once you get used to them they are as easy as any other and cheaper too.
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fngbronco
Monkey Man



Registered: 09/26/10
Posts: 2,877
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Re: Losing the battle with bacrteria [Re: Doc_T]
#14023323 - 02/24/11 11:51 PM (13 years, 8 days ago) |
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It's clean in pf teks but the bacteria gets my grain jars before they colonize.
-------------------- I challenge you to challenge yourself more! When you feel complacent and ready to hang it up, challenge yourself to get over it! If you fail, don't look at it as you didn't succeed, look at it as you would a rock face you're trying to climb. Stand back, wayyyy back, and look at it and plot another path. If you can't find one, shuffle down the way a little, a little change of scenery or a view from a different angle may give you the insight you need. Anything I state is relayed information from a friend of a friend and should be viewed as completely fictitious. I do not partake in any illegal or grey-area-of-the-law activities, but do have lots of friends who may or may not. -fngbronco Pill Divider Agar Tek
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heeroyuy00
Sublime


Registered: 09/27/09
Posts: 299
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Re: Losing the battle with bacrteria [Re: fngbronco]
#14024104 - 02/25/11 06:13 AM (13 years, 8 days ago) |
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yeah but you said your jar colonized to 75% then stopped? and how do you know they stink? your not opening the jars are you ?
-------------------- "You ever been to a day care center? Its like a fuckin' frat house... everyones sleeping all over the floor and all the girls are running around pulling there shirts up and tryna put everything in there mouth." Good Links
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Doc_T
Random Dude




Registered: 03/06/09
Posts: 42,395
Loc: Colorado
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Re: Losing the battle with bacrteria [Re: fngbronco]
#14024232 - 02/25/11 07:33 AM (13 years, 8 days ago) |
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Quote:
fngbronco said: It's clean in pf teks but the bacteria gets my grain jars before they colonize.
And your agar plates.
-------------------- You make it all possible. Doesn't it feel good?
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fngbronco
Monkey Man



Registered: 09/26/10
Posts: 2,877
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Re: Losing the battle with bacrteria [Re: Doc_T]
#14025424 - 02/25/11 01:06 PM (13 years, 8 days ago) |
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I've got Tyvek lids on half and the others you can smell through the nail holes. They also have that slimy shit that sticks to the glass.
-------------------- I challenge you to challenge yourself more! When you feel complacent and ready to hang it up, challenge yourself to get over it! If you fail, don't look at it as you didn't succeed, look at it as you would a rock face you're trying to climb. Stand back, wayyyy back, and look at it and plot another path. If you can't find one, shuffle down the way a little, a little change of scenery or a view from a different angle may give you the insight you need. Anything I state is relayed information from a friend of a friend and should be viewed as completely fictitious. I do not partake in any illegal or grey-area-of-the-law activities, but do have lots of friends who may or may not. -fngbronco Pill Divider Agar Tek
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Doc_T
Random Dude




Registered: 03/06/09
Posts: 42,395
Loc: Colorado
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Re: Losing the battle with bacrteria [Re: fngbronco]
#14025466 - 02/25/11 01:19 PM (13 years, 8 days ago) |
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Tyvek is not my favorite filter material. As part of changing one thing at a time to find the problem, grab a bag of polyfil and try it, see what you think. But I don't think that's the problem.
-------------------- You make it all possible. Doesn't it feel good?
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fngbronco
Monkey Man



Registered: 09/26/10
Posts: 2,877
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Re: Losing the battle with bacrteria [Re: Doc_T]
#14025691 - 02/25/11 02:12 PM (13 years, 8 days ago) |
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I'll set up the polyfil lids for sure. Heading to the store now!
-------------------- I challenge you to challenge yourself more! When you feel complacent and ready to hang it up, challenge yourself to get over it! If you fail, don't look at it as you didn't succeed, look at it as you would a rock face you're trying to climb. Stand back, wayyyy back, and look at it and plot another path. If you can't find one, shuffle down the way a little, a little change of scenery or a view from a different angle may give you the insight you need. Anything I state is relayed information from a friend of a friend and should be viewed as completely fictitious. I do not partake in any illegal or grey-area-of-the-law activities, but do have lots of friends who may or may not. -fngbronco Pill Divider Agar Tek
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