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Thomsen



Registered: 09/27/19
Posts: 364
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APE on agar - Da fuq is dis
#26927811 - 09/10/20 08:10 AM (3 years, 4 months ago) |
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Hey guys.
I'm cleaning APE on agar and it's acting up. I've made a total of 4 transfers from the first inoc and the first two transfers (multiple plates) looked fine, and on the 3rd and 4th transfer it looked like this in three plates out of 5.
It looks pretty odd and i've done 20 other plates at the same time i did these APE's and none of them are showing the same morphology. These pictures are of the 4th transfer and just like the 3rd transfer it seems to grow mostly alongside the edge of the dish.

The dishes is double wrapped with triple folded parafilm...
Any ideas?
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meowjinx
Stranger

Registered: 05/20/19
Posts: 492
Last seen: 1 year, 6 months
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Re: APE on agar - Da fuq is dis [Re: Thomsen]
#26927823 - 09/10/20 08:24 AM (3 years, 4 months ago) |
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Quote:
Thomsen said: Hey guys.
I'm cleaning APE on agar and it's acting up. I've made a total of 4 transfers from the first inoc and the first two transfers (multiple plates) looked fine, and on the 3rd and 4th transfer it looked like this in three plates out of 5.
It looks pretty odd and i've done 20 other plates at the same time i did these APE's and none of them are showing the same morphology. These pictures are of the 4th transfer and just like the 3rd transfer it seems to grow mostly alongside the edge of the dish.

The dishes is double wrapped with triple folded parafilm...
Any ideas?
Do you transfer multiple pieces onto one plate? Because (I'm sure you know this alread) it shouldn't be growing that patchy after 4 transfers. Those look like spore syringe plates
And did you fumble the transfer wedge? Cause you see almost like little satellite colonies around the 4 main blotches
Overall, I'd say you should go back to your previous generation of plates if you still have them and try to re-do the transfer. That whole plate looks way too suspect
Edited by meowjinx (09/10/20 08:25 AM)
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Thomsen



Registered: 09/27/19
Posts: 364
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Re: APE on agar - Da fuq is dis [Re: meowjinx]
#26927838 - 09/10/20 08:38 AM (3 years, 4 months ago) |
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Yeah i did 4 transfers onto one plate. These are the inoc points.

No fumbeling with the wedges when wrapped. Yeah the satelites looks super weird. I've seen it when i've cut into the agar to get the myc off the scapel but nothing like this.
This is two different plates btw.
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redhandmat
Dude


Registered: 05/09/19
Posts: 463
Last seen: 2 years, 8 months
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Re: APE on agar - Da fuq is dis [Re: Thomsen]
#26927857 - 09/10/20 08:56 AM (3 years, 4 months ago) |
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Yeah this is very weird. Have you pretended to inoculate a plate with nothing?
Like flame scalpel, touch it here and there on the agar, close the dish, wrap it and let it sit for a couple weeks. If your procedure is good then nothing should grow on it. If you see anything growing then something with the inoculation or pre-inoculation process is not right.
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Thomsen



Registered: 09/27/19
Posts: 364
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Re: APE on agar - Da fuq is dis [Re: redhandmat]
#26927860 - 09/10/20 08:58 AM (3 years, 4 months ago) |
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Yeah I do NTC's with every batch of agar. All clear so far
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redhandmat
Dude


Registered: 05/09/19
Posts: 463
Last seen: 2 years, 8 months
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Re: APE on agar - Da fuq is dis [Re: Thomsen]
#26927885 - 09/10/20 09:19 AM (3 years, 4 months ago) |
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Have you considered starting from spore plate or even from syringe again and do it all over?
Or maybe even better, sandwitch it with a hotpour and then transfer the growth to a new plate.
And do one transfer per plate. The growth looks a bit crazy for 4th transfer honestly.
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Thomsen



Registered: 09/27/19
Posts: 364
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Re: APE on agar - Da fuq is dis [Re: redhandmat]
#26928234 - 09/10/20 12:55 PM (3 years, 4 months ago) |
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I have and i do have other plates with APE that looks fine, also with single inocs. I'm just curious about what this could be. It kinda looks like areal spreaded myc.
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sporecap
Shedding...

Registered: 07/30/18
Posts: 413
Loc: Europe
Last seen: 1 year, 10 months
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Re: APE on agar - Da fuq is dis [Re: Thomsen]
#26928272 - 09/10/20 01:12 PM (3 years, 4 months ago) |
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I'd say everything besides the myc around your transfer wedges is some kind of mold contamination. Being located directly and the edge of the dish is usually a good indicator, and you can see at least at these points here that the cube myc and the other myc avoid each other and don't want to fuse as soon as they touch:
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