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Offlinebiohazard-unicorn
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Registered: 09/13/16
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Rookie needs assistance!! Possible contamination
    #23640588 - 09/13/16 04:23 AM (7 years, 4 months ago)

Hello to anyone reading, first off this is completely new to me.A few of these jars have wbs and a few jars with an oat mixture that I have a few questions about possible contamination. They were inoculated rough a week,w eek and a half ago and was wondering if it looks like just excessive water or if it would be some sort of contamination. Thank you in advance for taking a look.   





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OfflineTira
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Registered: 11/20/10
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Re: Rookie needs assistance!! Possible contamination [Re: biohazard-unicorn]
    #23640611 - 09/13/16 04:43 AM (7 years, 4 months ago)

That is definitely bacteria.


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Offlinedeph
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Re: Rookie needs assistance!! Possible contamination [Re: Tira]
    #23640620 - 09/13/16 04:52 AM (7 years, 4 months ago)

yeah not looking too good id toss, definitely too wet


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OfflineTira
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Re: Rookie needs assistance!! Possible contamination [Re: deph]
    #23640631 - 09/13/16 05:06 AM (7 years, 4 months ago)

That wet look is caused by bacteria.  Those jars are colonised by bacteria. Excessive water may or may not be the problem.


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Offlinebiohazard-unicorn
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Re: Rookie needs assistance!! Possible contamination [Re: Tira]
    #23641223 - 09/13/16 10:53 AM (7 years, 4 months ago)

Thank you for taking a look!


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OfflineTira
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Re: Rookie needs assistance!! Possible contamination [Re: biohazard-unicorn]
    #23641322 - 09/13/16 11:45 AM (7 years, 4 months ago)

No problem at all, but those jars look terrible. What did you inoculate those with?  How long did you PC them?  What kind of lids do you have on them?


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Offlinebiohazard-unicorn
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Re: Rookie needs assistance!! Possible contamination [Re: Tira]
    #23643413 - 09/13/16 11:30 PM (7 years, 4 months ago)

I inoculated with roughly 2.5 cc's out of spore syringes from sporestore.com I have read more than a few pretty bad reviews about that company(after purchase unfortunately) I have regular mason jar lids with two small nail holes poked in the top. I had pc'd for approximately 45 minutes. I did a little more reading and it seems most people do it for an hour to an hour and a half? Also I did a little more research and some people are saying grains may not be the best to start with for a beginner?


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InvisibleMad Season
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Re: Rookie needs assistance!! Possible contamination [Re: biohazard-unicorn]
    #23643488 - 09/13/16 11:57 PM (7 years, 4 months ago)

I do 2 hr pc cycles lol, and if it doesn't get colonized by a month or so, bacterial endospores start going crazy. Fuckin endospores..

The people who said that are dumb. I started off doing grains. I have no regrets. Also I'd recommend getting into agar for grains. It'll teach you epic sterile techniques in a still air box, and you can isolate out clean shit, even when in the presence of molds and Bacteria, because of the fact it's a 2d surface.


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Offlinebiohazard-unicorn
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Re: Rookie needs assistance!! Possible contamination [Re: Mad Season]
    #23643623 - 09/14/16 01:24 AM (7 years, 4 months ago)

I will restart and definitely give it more Time in the PC seeing as contamination is my problem!! I have seen a few people mention agar but haven't really looked into it but I will do some definitely give it an in depth read. Thank you for the knowledge assistance! Out of curiosity do you you have a preference on spore syringes or prints? Do you have any recommendations on a good company to get them from?


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OfflineTira
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Re: Rookie needs assistance!! Possible contamination [Re: Mad Season]
    #23647441 - 09/15/16 12:51 PM (7 years, 4 months ago)

Quote:

biohazard-unicorn said:
I had pc'd for approximately 45 minutes. I did a little more reading and it seems most people do it for an hour to an hour and a half? Also I did a little more research and some people are saying grains may not be the best to start with for a beginner?




Should PC longer, at least 90 minutes, guess best is to do it for 2 hours to be safe.

and PF tek is a safe way for a beginner to go but agar and grains are the real shit if you want to get serious.  Maybe do them both together? Do some cakes and do some grains at the same time, it'll be a good experience for you.

Quote:

Mad Season said:
I do 2 hr pc cycles lol, and if it doesn't get colonized by a month or so, bacterial endospores start going crazy. Fuckin endospores..




Even soaking and PCing for 2 hours after that does not kill endospores huh?


--------------------
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The Basics              AMU Teks

Frank''s Teks          Agar

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Agar for guaranteed spawn,
Proper pasteurization for guaranteed substrate.


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Offlinekatbusa
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Re: Rookie needs assistance!! Possible contamination [Re: Tira]
    #23657291 - 09/18/16 10:57 PM (7 years, 4 months ago)

Any of the vendors here are good. My personal favorite is sporeworks. Been using them for years and they have never failed me. They always send me good stuff.

I PC just about everything at 90min or more with the exception of agar which gets 25min.

Agar is probably one of the best ways to practice your work flow. Its easy to do but very unforgiving when it comes to contamination. If you can make clean agar plates your work flow is good.

Most of all its your methodology on how you work with things that are sterilized. You are the number one vector and with each failure you will adopt new methods. For example never working directly over top of sterilized material. Only removing lids to inoculate and replacing them immediately after. Never working upstream in a flow hood.

You can not become proficient if you don't experience failures. Live and learn! You will get the hang of it.


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Offlinebiohazard-unicorn
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Re: Rookie needs assistance!! Possible contamination [Re: Tira]
    #23657656 - 09/19/16 03:21 AM (7 years, 4 months ago)

Thank you very much for the input! I will definitely PC longer next time! I may take up the recommendation to try a little of both, see what works best for me. It will be a little while before I can restart due to unfortunate circumstances but I would love to come back with some positive results!! Again thank you for sharing the knowledge it is most appreciated :smile:


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Offlinebiohazard-unicorn
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Re: Rookie needs assistance!! Possible contamination [Re: katbusa]
    #23657663 - 09/19/16 03:26 AM (7 years, 4 months ago)

Thank you for the recommendation I will defintly be taking some more notes and trying to improve! Thank you for the knowledge and support.


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