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OfflineLSDevious
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Registered: 07/18/11
Posts: 18
Last seen: 9 years, 5 months
decontaminating agar
    #20350619 - 07/30/14 11:08 AM (9 years, 6 months ago)

So I'm a beggi er at all this  trying to decontaminate potatoes dextrose agar that I cultivated with spores from a dry mushroom cap I've got I was wondering if anyone one can give me a step by step/tips and valid information I may need all info will be greatly appreciated


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OfflineFivel
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Registered: 07/09/14
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Loc: Midwest
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Re: decontaminating agar [Re: LSDevious]
    #20350647 - 07/30/14 11:18 AM (9 years, 6 months ago)



--------------------
"The only way to deal with an unfree world is to become so absolutely free that your very EXISTENCE is an act of rebellion."
- Albert Camus  :bonghit:

Have Orissa Prints for trade!

In search of PF albino, PF redspore, PF classic :pm:

"... the associations between sadism and GAIT (Global Assessment of Internet Trolling) scores were so strong that it might be said that online trolls are prototypical everyday sadists."

:mushroom2::mushroom2::mushroom2::mushroom2::mushroom2::meltdown::mushroom2::mushroom2::mushroom2::mushroom2::mushroom2:


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OfflineTryptoman
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Registered: 06/13/13
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Re: decontaminating agar [Re: LSDevious]
    #20350665 - 07/30/14 11:24 AM (9 years, 6 months ago)

I am assuming that you have myc growing on these plates and that you also have some other nasties growing too.  Now this is more from experience working in a molecular biology lab than anything else, but if you cut out (use a McDicks straw sterilized with alcohol) and place the exert on a new plate and continue growing, you can potentially select against any contaminants present in the original plate.  I would take multiple sub-samples from the original plate and give each exert their own plate to grow on. By doing this you increase the probability that at least one of those exerts do not contain a con tam (use a new straw every time).  Repeat as necessary. 

Happy Growing


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OfflineMdahmer
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Re: decontaminating agar [Re: Tryptoman]
    #20350751 - 07/30/14 11:51 AM (9 years, 6 months ago)

For agar transfers you need to use a metal tool like a scalpel or thin metal that can be flame sterilized (red hot) and used to cut wedges of agar. Alcohol only sanitizes and I can't imagine a straw being very convenient for transfers


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OfflineSockadin
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Re: decontaminating agar [Re: Mdahmer]
    #20350833 - 07/30/14 12:17 PM (9 years, 6 months ago)

The straw seems like a great idea. I brb gotta go find a metal straw.


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OfflineTryptoman
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Re: decontaminating agar [Re: Mdahmer]
    #20350855 - 07/30/14 12:24 PM (9 years, 6 months ago)

I know it sounds fairly sketchy but I have seen those straws survive the autoclave before, just toss em in a jar with some foil over and let em go.  Best case scenario you have autoclaved straws... worst case you have a bunch of melted plastic that you can mould into beautiful desktop art!


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OfflineLSDevious
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Re: decontaminating agar [Re: Tryptoman]
    #20351020 - 07/30/14 01:28 PM (9 years, 6 months ago)

Trypto what do u mean by mcdicks straw


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InvisibleEskeer
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Registered: 06/23/14
Posts: 110
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Re: decontaminating agar [Re: Tryptoman]
    #20351027 - 07/30/14 01:31 PM (9 years, 6 months ago)

how do you get the piece of agar in the straw out of it onto the next plate? This is something thats been bothering me in my transfers, the piece of agar often gets stuck to my scalpel (especially if I stab it to get it on the scalpel) causing the new plate to be exposed to air for far longer than I would like(while I wriggle the scalpel around to get the piece off).

mcdicks = mcdonalds :tongue2:


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Feel free to comment/suggest/moan.


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OfflineLSDevious
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Re: decontaminating agar [Re: Eskeer]
    #20351036 - 07/30/14 01:34 PM (9 years, 6 months ago)

Thanks eskeer I thought that but I jus wanted to be sure


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OfflineFivel
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Re: decontaminating agar [Re: LSDevious]
    #20351131 - 07/30/14 01:57 PM (9 years, 6 months ago)

Use an exacto knife. They are as low as 3 dollars at some places. Flame sterilization is pretty sure fire.


--------------------
"The only way to deal with an unfree world is to become so absolutely free that your very EXISTENCE is an act of rebellion."
- Albert Camus  :bonghit:

Have Orissa Prints for trade!

In search of PF albino, PF redspore, PF classic :pm:

"... the associations between sadism and GAIT (Global Assessment of Internet Trolling) scores were so strong that it might be said that online trolls are prototypical everyday sadists."

:mushroom2::mushroom2::mushroom2::mushroom2::mushroom2::meltdown::mushroom2::mushroom2::mushroom2::mushroom2::mushroom2:


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Offlinetombosley8
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Registered: 10/14/13
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Re: decontaminating agar [Re: Fivel]
    #20351599 - 07/30/14 03:35 PM (9 years, 6 months ago)

:whathesaid:


--------------------


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OfflineSockadin
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Registered: 01/03/10
Posts: 7,244
Last seen: 3 months, 7 days
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Re: decontaminating agar [Re: tombosley8]
    #20352927 - 07/30/14 08:21 PM (9 years, 6 months ago)

Im gonna try the metal straws. Google Metal straws and you can buy them cheep. Then you can flame sterilize. This is all experimental and I don't recommend it to anyone.


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