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ProteinDrink
Apprentice


Registered: 07/17/23
Posts: 48
Loc: State of Confusion
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Why does my APE keep contaminating?
#28464245 - 09/10/23 11:27 AM (1 year, 4 months ago) |
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I have done a couple of successful grows, and have successfully colonized four different strains in WBS.
However, I have also tried colonizing APE twice in the same manner as the four other successful strains, and both times the jars started to colonize, and wherever mycelium started to grow, contams took over : (
I assumed that the inoculant was conatmmed, but I have transferred 3 times on agar, and each time the sample looks good, yet once introduced to my jars, failure : (
I have done yet another transfer. I also took a shot at grabbing some of the seeds from one of the contammed jars, as far from contam as possible, and tried growing that out on agar.
Here are pics of two 'plates.' One is from a transfer of what I thought was clean growth on agar > agar > this third plate, and the other is a pieces of the grain which has grown out.

Both look good to me, yet I expect they will fail again on my third try.
Your thoughts on this culture?
Does the APE need a different recipe for the jars (different grain, different moisture, additives, etc.)?
Thanks for any input. I really want to grow some APE, and it bums me out to keep making jars and then tossing them : (
-------------------- The comments above are made up, and are only intended for amusement. All photos have been gathered from the internet, and all content is fictional.
"If the thunder don't get you, then the lightning will."
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Screwup
Googles your dumb questions


Registered: 01/27/22
Posts: 6,340
Last seen: 10 months, 7 days
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Re: Why does my APE keep contaminating? [Re: ProteinDrink] 1
#28464248 - 09/10/23 11:29 AM (1 year, 4 months ago) |
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I canβt see shit
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stareatclouds
star eat clouds?



Registered: 09/29/14
Posts: 10,361
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Re: Why does my APE keep contaminating? [Re: Screwup]
#28464268 - 09/10/23 11:48 AM (1 year, 4 months ago) |
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They look like cotton swabs, man. No, those don't look clean to me. Even if they are, you should try and get flatter, more observable growth with your transfers. The benefit of agar is it's a flat, 2D surface that you can observe growth on easier. Those jars are already more difficult to see into due to the glass. Plopping in grains or huge chunks of agar aren't conducive to flat, 2D growth.
Don't put grains to agar anyway, let alone contaminated ones. Streak spores or use a few drops from a syringe. When you're newer, starting from as clean of a source possible is best.
     
I'm not saying all of those were clean, either. The left pic with PPs are all clones. Tissue samples always puff up like that for me and spread out. In the center where it's puffed, I have no idea wtf is in there. So I wait for it to smooth out so I can actually observe the health of the myc.
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ProteinDrink
Apprentice


Registered: 07/17/23
Posts: 48
Loc: State of Confusion
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Re: Why does my APE keep contaminating? [Re: stareatclouds]
#28464280 - 09/10/23 11:59 AM (1 year, 4 months ago) |
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Quote:
stareatclouds said: They look like cotton swabs, man. No, those don't look clean to me. Even if they are, you should try and get flatter, more observable growth with your transfers. The benefit of agar is it's a flat, 2D surface that you can observe growth on easier. Those jars are already more difficult to see into due to the glass. Plopping in grains or huge chunks of agar aren't conducive to flat, 2D growth.
Don't put grains to agar anyway, let alone contaminated ones. Streak spores or use a few drops from a syringe. When you're newer, starting from as clean of a source possible is best.
I only did the grain plate to see what it would do. What frustrates me is that even the grain on the agar is not contammed (yet), but every jar ultimately had all the white taken over by grey : (
What you shared about the flat growth makes perfect sense. That is good info. I can use. Instead of agar to agar by way of 'chunks,' should I be just touching a needle to the mycelia and then streak with that? I assume I don't have to actually see white to successfully seed a plate?
I am not happy with the small jars (though they are a small step up from disposable food containers). I am going to get some actual petri dishes for my next cultures. Probably get a spore syringe as well, and start from scratch.
Thanks for the helpful feedback.
-------------------- The comments above are made up, and are only intended for amusement. All photos have been gathered from the internet, and all content is fictional.
"If the thunder don't get you, then the lightning will."
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Kinoko314
Stranger Danger



Registered: 12/16/22
Posts: 1,585
Loc: Colorado
Last seen: 4 months, 1 day
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Re: Why does my APE keep contaminating? [Re: ProteinDrink]
#28464306 - 09/10/23 12:22 PM (1 year, 4 months ago) |
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It looks pretty disorganized, and it's puffed up a bit. Both can indicate that it's not clean. There is probably bacteria riding along.
You could take a transfer to water agar, then back to regular agar.
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AspectOfTheCreator
Mastering the Art of Success



Registered: 12/07/22
Posts: 2,179
Last seen: 8 hours, 51 minutes
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Re: Why does my APE keep contaminating? [Re: ProteinDrink]
#28464311 - 09/10/23 12:27 PM (1 year, 4 months ago) |
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Quote:
ProteinDrink said:
Quote:
stareatclouds said: They look like cotton swabs, man. No, those don't look clean to me. Even if they are, you should try and get flatter, more observable growth with your transfers. The benefit of agar is it's a flat, 2D surface that you can observe growth on easier. Those jars are already more difficult to see into due to the glass. Plopping in grains or huge chunks of agar aren't conducive to flat, 2D growth.
Don't put grains to agar anyway, let alone contaminated ones. Streak spores or use a few drops from a syringe. When you're newer, starting from as clean of a source possible is best.
I only did the grain plate to see what it would do. What frustrates me is that even the grain on the agar is not contammed (yet), but every jar ultimately had all the white taken over by grey : (
What you shared about the flat growth makes perfect sense. That is good info. I can use. Instead of agar to agar by way of 'chunks,' should I be just touching a needle to the mycelia and then streak with that? I assume I don't have to actually see white to successfully seed a plate?
I am not happy with the small jars (though they are a small step up from disposable food containers). I am going to get some actual petri dishes for my next cultures. Probably get a spore syringe as well, and start from scratch.
Thanks for the helpful feedback.
People wont like hearing this but no pours suck. I wasted months trying to get them to work. Went from 90% failure rate to 0% practically overnight once I swapped to pouring traditional agar plates
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ProteinDrink
Apprentice


Registered: 07/17/23
Posts: 48
Loc: State of Confusion
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Re: Why does my APE keep contaminating? [Re: Kinoko314]
#28464313 - 09/10/23 12:28 PM (1 year, 4 months ago) |
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Quote:
Kinoko314 said: It looks pretty disorganized, and it's puffed up a bit. Both can indicate that it's not clean. There is probably bacteria riding along.
You could take a transfer to water agar, then back to regular agar.
Interesting. I didn't realize that nice fluffy white growth could be harboring a hitch-hiker.
I had not seen anything about water agar, but a quick search led to this:
https://www.shroomery.org/forums/showflat.php/Number/27900393
I will try this.
Thanks for the tip.
-------------------- The comments above are made up, and are only intended for amusement. All photos have been gathered from the internet, and all content is fictional.
"If the thunder don't get you, then the lightning will."
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Screwup
Googles your dumb questions


Registered: 01/27/22
Posts: 6,340
Last seen: 10 months, 7 days
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I feel like youβre the exception not the rule. My no pours are practically flawless.
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π
π° πΏ
Help US help YOU TEK
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