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Author Topic: A major f**up?  (Read 2505 times)

spp.

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A major f**up?
« on: April 10, 2010, 12:31:18 PM »


So there, today I was inoculating some PF jars when the syringe needle detached. I immediately wiped it with H202, sticked it back and continued inoculating tha jars, but now I'm worried if it wasn't enough. Basically I was working in a glowbox chiefly sprayed with peroxide, but hey shit happened and I'm afraid all my jars are doomed now. Has anyone had such an experience?
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MT

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Re: A major f**up?
« Reply #1 on: April 12, 2010, 09:41:40 PM »

If you were working with a glove box and everthing was already sterile, you should be okay.
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spp.

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Re: A major f**up?
« Reply #2 on: April 13, 2010, 10:17:53 AM »

Well I'm not absolutely sure it was sterile, because I only used peroxide. Time will show. For now, it's been 3 days after inoculation and in every single jar the mycelium has made a colonization spot at least 2cm in diameter. If there are contaminants I think they will have shown up by the end of the week at 30 Celsius. Will inform you on how it's going.

By the way, as for sterility, currently I'm having microbiology classes, which have shown that, well, the common sterilizing agents we use pretty much suck. Hands cleaned with 3 agents [subsequently][/b]- antibacterial soap, 98% alcohol, and biguanide (antibacterial desinfectant) still had nice amount of various spores which grew into pretty colourful colonies on agar plates.
Given all that, I really doubt I was working sterile  -_- . Results have also shown that alcohol is the weakest desinfectant possible, 3% peroxide being a little better. 2% jodine solution is the best from the easily obtainable ones, but its use in mycology will be probably limited due to heavy staining and it being pretty unhealthy to breath.
« Last Edit: April 13, 2010, 10:35:23 AM by spp. »
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jeff_diggler

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Re: A major f**up?
« Reply #3 on: April 14, 2010, 10:18:58 AM »

try lysol to sterilize your glove box and 70% alcohol to clean it, peroxide isnt enough, if its clean you should be good on your needle problem. what i do to make sure the needle is on is give it a twist as you pull the cap off, do it enough times and you wont even have to think about it, just get in the habit of twisting it snug as you remove the cap. ive had several loose needles right out of the package, you'd be surprised.
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olzo

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Re: A major f**up?
« Reply #4 on: April 14, 2010, 10:43:35 AM »

Sorry guys but this is a bit ridiculous. You're all right when it comes to inoculating petri dish. Using syringes and burning the tip red hot in between jars will perfectly do the job with PF-Tek. Last time I've inoculated it wasn't even in a sterile environment yet no contamination happened. So chill a bit, use the syringes and burn the tip in between jars... tellin ya it will be alright.  -_-
« Last Edit: April 14, 2010, 10:45:21 AM by olzo »
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spp.

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Re: A major f**up?
« Reply #5 on: April 14, 2010, 11:52:08 AM »

Will see  -_- Gonna check'em on Saturday. I suppose if they are clear 7 days after inoculation I was lucky enough  :rolleyes:
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