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Offlinemushhiehunter
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guess the contam rate game
    #23745377 - 10/17/16 11:55 AM (7 years, 3 months ago)

Ok guys, so I had some extra agar jars and some extra time on my hands so I decided to do everything wrong and see what the contam rate is...

procedure aka "how it's not supposed to be done":
I poured the agar into jars, let them open till complete cool down in a kitchen with an open window then I PCd them for 25 minutes, then let the pressure out quickly and took away the Al foil (having tyvek/silicone injection point and AE) then I put a drop of bleach on each inj. point and let them cool to room temp and inoculated without glovebox sterilizing the needle only for first two jars :laugh:

So I have 6 agar plates

What are your guesses for contam rate? :laugh:
PS: will update when the germination/contamination begins...

Please take this only as a "anectdotal experiment"


Edited by mushhiehunter (10/17/16 11:56 AM)


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OfflinePortabellaFella 1
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Re: guess the contam rate game [Re: mushhiehunter]
    #23745405 - 10/17/16 12:10 PM (7 years, 3 months ago)

I guess 100%


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Invisibleblindingleaf
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Re: guess the contam rate game [Re: PortabellaFella 1]
    #23745418 - 10/17/16 12:14 PM (7 years, 3 months ago)

Letting them cool in open dirty air isn't really a vector.  Ur pcing them after.

If u used an injection port I feel like ur contam rate is gonna be more affected by the spore syringe than anything else.

I guess either all 6 contam or all 6 stay clean :lol:


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Invisibleamidogen
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Re: guess the contam rate game [Re: blindingleaf]
    #23745428 - 10/17/16 12:17 PM (7 years, 3 months ago)

.


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The biggest trip of my life was realizing all of the events and actions described in posts made by this account were never real and had never actually happened, but were instead the delusional ramblings of a severely mentally ill human being. I just had to move on for my own good. I love you all.


Edited by amidogen (04/19/18 11:44 AM)


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InvisibleJosex
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Re: guess the contam rate game [Re: PortabellaFella 1]
    #23745429 - 10/17/16 12:17 PM (7 years, 3 months ago)

There's nothing to guess here, those are gonna be nasty  :studmuffin:

Is this another "I'm telling people that I'm doing an experiment to see if I can get away without doing proper sterile technique" thread?


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Offlinemushhiehunter
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Re: guess the contam rate game [Re: Josex]
    #23745435 - 10/17/16 12:20 PM (7 years, 3 months ago)

Quote:

Josex said:
There's nothing to guess here, those are gonna be nasty  :studmuffin:

Is this another "I'm telling people that I'm doing an experiment to see if I can get away without doing proper sterile technique" thread?




Not really, I just have 20 agar jars going which were prepared correctly so I just want to see what will be the difference between the "control" sample and the "dirty redneck" one... :laugh:


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InvisibleJosex
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Re: guess the contam rate game [Re: mushhiehunter]
    #23745443 - 10/17/16 12:25 PM (7 years, 3 months ago)

Nah... you just don't wanna do sterile tech :awehigh:


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Offlinemushhiehunter
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Re: guess the contam rate game [Re: blindingleaf]
    #23745503 - 10/17/16 12:46 PM (7 years, 3 months ago)

Quote:

blindingleaf said:
Letting them cool in open dirty air isn't really a vector.  Ur pcing them after.

If u used an injection port I feel like ur contam rate is gonna be more affected by the spore syringe than anything else.

I guess either all 6 contam or all 6 stay clean :lol:





The dirty air is a contam vector because the more contams get in the more have to be killed by PCing. If the agar after pouring had 1000 contam spores on it the chance of them dying during the 25 minutes @121C is much higher than if there is 1 000 000 spores :smile:


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InvisibleJosex
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Re: guess the contam rate game [Re: mushhiehunter]
    #23745529 - 10/17/16 12:53 PM (7 years, 3 months ago)

No matter the initial spore load in your plates, all the spores are going to be destroyed during the pc run, spores will die at temperatures below boiling point.
It's bacteria that's harder to kill, but in a liquid media like agar it's also easy to kill all the bacteria present in the plates during pc'ing.


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Offlinemushhiehunter
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Re: guess the contam rate game [Re: Josex]
    #23745533 - 10/17/16 12:55 PM (7 years, 3 months ago)

Quote:

Josex said:
No matter the initial spore load in your plates, all the spores are going to be destroyed during the pc run, spores will die at temperatures below boiling point.
It's bacteria that's harder to kill, but in a liquid media like agar it's also easy to kill all the bacteria present in the plates during pc'ing.




yeah makes sense, thanks for an insight!


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OfflinePortabellaFella 1
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Re: guess the contam rate game [Re: mushhiehunter]
    #23745535 - 10/17/16 12:56 PM (7 years, 3 months ago)

The whole point of PCing is to kill off the contams acquired pre PC. If that didn't work then no ones agar would work.


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Offlinethebug76
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Re: guess the contam rate game [Re: PortabellaFella 1]
    #23745693 - 10/17/16 02:02 PM (7 years, 3 months ago)

I'm going to guess 4 contaminated and the 2 that you flamed the needle will be clean.


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Offlinemushhiehunter
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Re: guess the contam rate game [Re: thebug76]
    #23745716 - 10/17/16 02:07 PM (7 years, 3 months ago)

Quote:

thebug76 said:
I'm going to guess 4 contaminated and the 2 that you flamed the needle will be clean.




I have those two marked so we'll see... it's funny how I am more interested in the results of those made the crude way than the "clean" ones..


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OfflinePlazmotech
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Re: guess the contam rate game [Re: mushhiehunter]
    #23746055 - 10/17/16 03:27 PM (7 years, 3 months ago)

2/6


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OfflineMysticMoteToter
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Re: guess the contam rate game [Re: Plazmotech]
    #23746281 - 10/17/16 04:29 PM (7 years, 3 months ago)

Quote:

mushhiehunter said:...it's funny how I am more interested in the results of those made the crude way than the "clean" ones..




thats some shit some bad milk would say, gonna get moldy dude!


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Offlinemushhiehunter
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Re: guess the contam rate game [Re: MysticMoteToter]
    #23764165 - 10/23/16 12:33 PM (7 years, 3 months ago)

Well guess what.. all of them are colonizing without signs of contamination :laugh: will supply images soon


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OfflinePortabellaFella 1
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Re: guess the contam rate game [Re: mushhiehunter]
    #23764175 - 10/23/16 12:37 PM (7 years, 3 months ago)

Quote:

mushhiehunter said:
Well guess what.. all of them are colonizing without signs of contamination :laugh: will supply images soon




I'd add "yet" after the word contamination... just for good measure :wink:


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Offlinemushhiehunter
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Re: guess the contam rate game [Re: PortabellaFella 1]
    #23764248 - 10/23/16 01:08 PM (7 years, 3 months ago)

Quote:

PortabellaFella 1 said:
Quote:

mushhiehunter said:
Well guess what.. all of them are colonizing without signs of contamination :laugh: will supply images soon




I'd add "yet" after the word contamination... just for good measure :wink:





I don't think the will have time to do that since the mycelium pretty much popped up over night and is colonizing quite quickly (yes I am sure it's mycelium) :laugh:


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Offlineomgwtfstfu
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Re: guess the contam rate game [Re: mushhiehunter]
    #23764334 - 10/23/16 01:51 PM (7 years, 3 months ago)

Not all contams are easy to spot by a novice (or even an expert). "Getting away" with doing it the wrong way only encourages bad habits. Luck does not equal good practice. Bragging about using bad technique will only encourage more noobs to try and fail. Doing things the right way does not ensure 100% success so why risk it even more? Nothing to be gained by this non-experiment.


Edited by omgwtfstfu (10/23/16 01:52 PM)


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Offlinemushhiehunter
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Re: guess the contam rate game [Re: omgwtfstfu]
    #23764342 - 10/23/16 01:54 PM (7 years, 3 months ago)

Quote:

omgwtfstfu said:
Not all contams are easy to spot by a novice (or even an expert). "Getting away" with doing it the wrong way only encourages bad habits. Luck does not equal good practice. Bragging about using bad technique will only encourage more noobs to try and fail. Doing things the right way does not ensure 100% success so why risk it even more? Nothing to be gained by this non-experiment.




Absolutely agree with you, I just couldn't keep my amazement to myself considering all the things that could (can still) have gone wrong...


Edited by mushhiehunter (10/23/16 01:55 PM)


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