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rexxxlo
Stranger

Registered: 03/26/06
Posts: 3
Last seen: 16 years, 11 months
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another noob contam ?
#5601135 - 05/07/06 02:03 PM (17 years, 8 months ago) |
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i have had 100% contam rate so far
its always the same thing black fuzz i even had it in some petri dishes a friend who works in a hospital got me that i never opened up they were contaminated right away i didn't even inoculate them i have tried the pf tek and got the same results so far except for the last 4 jars i have done seem to be almost fully colinated and looking good
here is a rundown of my procedures
1/2 pint ball jar 2 small nail holes
i found a used pressure cooker on ebay ( think this is the problem)
a glove box made from a sterilite clear tote holes cut in lid and large black rubber gloves duct tapped in to the lid lysol spray before i use it and a lighter to flame the needles
spore syringes from one of the sponsors on here 4 different varieties
an old Coleman cooler with a thermostat hooked to a light bulb cased in a piece of sheet metal to prevent light from escaping kept at 80-90 deg f
does it cause a problem if the light gets to them because i like to check on them to see how they look?
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mrTtheH
Micro Bio

Registered: 04/01/04
Posts: 188
Loc: C A N A D A
Last seen: 15 years, 4 months
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Re: another noob contam ? [Re: rexxxlo]
#5603003 - 05/07/06 10:26 PM (17 years, 8 months ago) |
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please indicate how long you are pressure cooking the jars for? You also stated that you purchased a used cooker. Do you think that you might not be getting a desired pressure? This isnt really that big of a problem. The pressure cooker that i own is in terrible shape, and i have very rarely have trouble with contamination. It very rarely gets a desired pressure but it boils and whistles. I like your usage of the glovebox. The usage of a glovebox in my opinion would cut down on 75% noobs getting contamination.
Also, one way to simply tell if your syringe is the source of the contamination is that the place where the liquid was transfered from the syringe from the jar will become contaminated quickly and be very obvious. You stated that you used two holes in the lid of the jar. Did the areas around the holes get contaminated first?
-------------------- It is the mark of an educated mind to be able to entertain a thought without accepting it. Aristotle (384 BC - 322 BC)
"Human beings, the potentially highest form of life expression on this planet have built the vast pharmaceutical industry for the central purpose of poisoning the lowest form of life on the planet--germs!"--Dr. Richard Murray
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rexxxlo
Stranger

Registered: 03/26/06
Posts: 3
Last seen: 16 years, 11 months
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Re: another noob contam ? [Re: mrTtheH]
#5613924 - 05/10/06 05:55 PM (17 years, 8 months ago) |
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well if its not mu pressure cooker and its not my syringe it must have been me,my glove box or my incubation cooler it must have been dirty i have sence cleaned it out again and inoculated 8 more pf jars with the 4 that were going still going strong i think they are about to be birthed soon what else could this have been i only see yellow green or white contamination problems on the forum not the black that i had ?
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MLBjammer
Invitro Freak


Registered: 02/24/06
Posts: 560
Loc: Southeastern USA
Last seen: 7 years, 14 days
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Re: another noob contam ? [Re: rexxxlo]
#5614243 - 05/10/06 06:56 PM (17 years, 8 months ago) |
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Contams will usually come from a bad or dirty syringe--or something already in the substrate. You shouldn't worry yourself to death. Just follow whatever tek you are using, use common sense in being clean especially during inoculation, and you will get the hang of it. It sounds as though you are having some success, anyway. GL.
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