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Invisiblebebop
Registered: 07/04/20
Posts: 125
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Agar germination speed
    #26924840 - 09/08/20 03:06 PM (3 years, 5 months ago)

Made some pastyplates and they've yet to germinate anything after 14 days, aside from one that contaminated very early on and was disposed of. I'm wondering if any of these details could be to blame for the lack of growth:

LME agar, with a little dextrose and peptone.

Inoculated from a spore print using disposable sterilized inoculation loops (one per plate). I streaked in a zig-zag. I can see the dull streak on each plate. Some of them I can see the black spores, one is a long black streak as the print was smudged and wet.

I initially blamed the print, but I inoculated some BRF cakes 6 days ago with an MS made from the same print, and 2 out of 3 cakes are showing some growth, though small. However, nothing on any of the agar plates.

I've been keeping the plates in a tub with the lid on top perpendicularly to leave openings, in a closet. Very little ambient light.

Temperatures move up and down but hover around high 70s.

Instead of drilling a hole in the top per Pasty's tek, I used a flathead screwdriver to make slits on the side covered in 2-3 layers of micropore tape. I would assume it's plenty for GFE but who knows?

I've seen people report it could take up to a month, but I'm wondering what factors into that most?


Edited by bebop (09/08/20 03:43 PM)


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OfflineFredsGotSlacks
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Registered: 02/11/20
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Re: Agar germination speed [Re: bebop]
    #26925021 - 09/08/20 05:19 PM (3 years, 5 months ago)

Nothing to add, just kinda the same situation for me, Pasty’s with the flathead on the side with MP tape for GE. Slow germination.
Wanting to read if anyone else has something to add.


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Offlinetryptkaloids
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Re: Agar germination speed [Re: FredsGotSlacks]
    #26925069 - 09/08/20 05:59 PM (3 years, 5 months ago)

Sorfter agar works wonders for spore germination
They need to be able to hydrate


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OfflineOne of Us
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Re: Agar germination speed [Re: tryptkaloids]
    #26925085 - 09/08/20 06:06 PM (3 years, 5 months ago)

^^^ this

Also, more nutes helps.  Not sure about potato flakes, but about 2% LME by weight is good for germination.

Spore age is also a big factor usually. But doesn't seem that's the issue in your case since your cakes worked.


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Invisiblebebop
Registered: 07/04/20
Posts: 125
Loc: nt.here_
Re: Agar germination speed [Re: One of Us]
    #26932420 - 09/12/20 08:02 PM (3 years, 5 months ago)

Tried a drop of MS from the same print on each of the plates. I'll give it a week. The cakes are taking off!

Good to know, though. I'll make sure it's softer the next batch


Edited by bebop (09/12/20 08:03 PM)


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