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OfflineChowder
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What did I do wrong? - [Easy Agar Tek]
    #20999101 - 12/19/14 02:14 PM (9 years, 3 months ago)

I used Pastywhyte's Easy Agar Tek fir my first ever agar try.

I did everything according to the recipe, except for using cotton swabs. Instead, I used sterile plastic ones. The whole procedure was done in my glove-box

The trouble is that all my plastic rounds got this type of contamination, including the control one, that didnt receive any spores. Close-up. It had a sweet smell.

What could have gone wrong? How can I improve my next attempt.

Also, can I use this tek for to isolate?


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InvisibleMr. Alien
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Re: What did I do wrong? - [Easy Agar Tek] [Re: Chowder]
    #20999115 - 12/19/14 02:19 PM (9 years, 3 months ago)

I blame your sterile procedures. You don't use Glove boxes for agar, you use a SAB. Yes you can use Pasty agar tek for cloning and isolating too.

That's bacteria. If the control one had the same luck than the others... Then the sterelization process failed. You didn't open the control container at the moment you was inoculating spores to the other pasty containers right?

Edited by Mr. Alien (12/19/14 02:33 PM)

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OfflineChowder
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Re: What did I do wrong? - [Easy Agar Tek] [Re: Mr. Alien]
    #20999481 - 12/19/14 04:03 PM (9 years, 3 months ago)

Sorry, I didnt know theres a difference between a glovebox and a SAB. Yes, it is the SAB that I used.

As I was writing the question, I did realize that I'm not sure whether the control was opened or not. I should take notes next time.

Is there any kind of hint the fact that its a bacterial contamination? Does it make it more likely that its the sterilization that failed, rather than the inoculation?

I guess I'll just have to try it again and again and write  everything down. Thanks for your input.


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InvisibleEnigma1
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Re: What did I do wrong? - [Easy Agar Tek] [Re: Chowder]
    #21000427 - 12/19/14 08:00 PM (9 years, 3 months ago)

Did You use a print or syringe?

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InvisiblebodhisattaMDiscordReddit
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Re: What did I do wrong? - [Easy Agar Tek] [Re: Enigma1]
    #21000439 - 12/19/14 08:03 PM (9 years, 3 months ago)

bacterial most of the time points to operator error but more rarely a bad source of inoculant(but even that is usually created by a bad operator IE noob print/syringes)

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OfflineChowder
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Re: What did I do wrong? - [Easy Agar Tek] [Re: bodhisatta]
    #21003578 - 12/20/14 03:03 PM (9 years, 3 months ago)

Yes, from print.
I do take lots of care with taking my prints. Un fanned room, in a plastic box, under jars, wipe foils with IPA beforehand etc etc


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InvisiblePastywhyteMDiscord
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Re: What did I do wrong? - [Easy Agar Tek] [Re: Chowder]
    #21003860 - 12/20/14 04:05 PM (9 years, 3 months ago)

If the control was not opened then its definitely too short of sterilization or too low of pressure or both. This seems likely.  How long did you PC and at what pressure?

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