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Offlinestraightslayer
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First agar mission
    #24798351 - 11/21/17 07:01 PM (20 days, 13 hours ago)

Hi guys,

My first time making agar and inoculating with spore print. Followed a TEK but obviously have some very obvious mold issues.

Couple of questions. The ones that have mold on them, should I throw them out now as they might potentially contaminate the plates which have not grown anything yet?

Also in the first photo with the plate nearest the bottom. Obviously a mold spot but what about the other cloud shaped growth? Is that mycelium? If so can I cut it out and transfer it to a new agar plate?



Thanks in advance


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OfflineWorkmanV
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Re: First agar mission [Re: straightslayer]
    #24800283 - 11/22/17 06:04 PM (19 days, 14 hours ago)

The moldy ones will over grow the entire plate before any good mycelium appears. It's best to toss those. The cloudy material is yeast or bacteria and may cover the entire plate, especially if condensation drips water on the agar and spreads it around.

In my experience, the white/clear bacterial slimes don't always kill the spores which may eventually germinate and grow beyond the perimeter of the slime. You can then isolate the clean edge of mycelium on a fresh plate. So don't automatically discard the slimy plates unless the whole plate is covered or there is also mold. You might get lucky.

Keep at it. Agar skills are well worth the effort.


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Reinvesting 25% of Sales Towards Basic Research and Species Identification :amanitajar:


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Offlinestraightslayer
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Re: First agar mission [Re: Workman]
    #24807741 - 11/26/17 03:48 PM (15 days, 16 hours ago)

Thanks a lot for your reply

I have some more recent photos. Am I looking any better? The white stuff mycelium or just bacteria?



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OfflineWorkmanV
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Re: First agar mission [Re: straightslayer]
    #24809437 - 11/27/17 12:44 PM (14 days, 19 hours ago)

All look to have mycelium. The one on the right looks best. The top left one with the mold spot also has a fast growing thin contaminate on the top edge. The bottom left one has a pinkish contaminate wedge.

All of these are ready to be transferred to fresh plates (although the moldy one is risky to open). The best looking mycelium is coming from the obvious spore streaks that can be seen through the mycelium.


--------------------
Research funded by the patrons of
The Spore Works
Exotic Spore Supply
Reinvesting 25% of Sales Towards Basic Research and Species Identification :amanitajar:


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Offlinestraightslayer
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Re: First agar mission [Re: Workman]
    #24810954 - 11/28/17 03:31 AM (14 days, 4 hours ago)

Thanks for your reply

I will transfer to fresh plates asap. I obviously just follow exact same principals with my SAB etc as when I initially inoculated them right?


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OfflineWorkmanV
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Re: First agar mission [Re: straightslayer]
    #24811482 - 11/28/17 01:06 PM (13 days, 19 hours ago)

Yes, assuming you are using proper sterile technique.


--------------------
Research funded by the patrons of
The Spore Works
Exotic Spore Supply
Reinvesting 25% of Sales Towards Basic Research and Species Identification :amanitajar:


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Offlinestraightslayer
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Re: First agar mission [Re: Workman]
    #24819573 - 12/01/17 07:05 PM (10 days, 13 hours ago)

Hi again,

Have an updated image of my transferred agar. Dishes are a couple of days old. Do they look ok so far? Excuse the dirty dish tops. I believe it's from my soap mix which I used in my SAB while my dishes were in there.




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Invisiblesomegirl
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Re: First agar mission [Re: straightslayer]
    #24820215 - 12/02/17 12:05 AM (10 days, 8 hours ago)

They look fine.  Next time try for smaller transfers, closer to the size of a grain of rice.


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Offlinestraightslayer
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Re: First agar mission [Re: somegirl]
    #24820523 - 12/02/17 05:40 AM (10 days, 2 hours ago)

Beauty, thank you and will do.


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OfflineWorkmanV
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Re: First agar mission [Re: straightslayer]
    #24825272 - 12/04/17 12:07 PM (7 days, 20 hours ago)

Yes, smaller transfers reduce the risk of transferring contamination and reduces the number of strains per plate (if that is important to you).


--------------------
Research funded by the patrons of
The Spore Works
Exotic Spore Supply
Reinvesting 25% of Sales Towards Basic Research and Species Identification :amanitajar:


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Offlinestraightslayer
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Re: First agar mission [Re: Workman]
    #24828954 - 12/05/17 11:37 PM (6 days, 8 hours ago)

Hi again,

Have some more recent photos and wanting some feedback. Exciting stuff and I think I have a few good looking dishes. Obviously a couple contams. The second photo actually originally did have a contam spot but the mycelium just grew straight over it and it's not even noticeable in this photo anymore. Would it still be ok to use that particular dish?

Also when can I use the clean dishes? Do I have to wait till the mycelium grows right out to the edges? I am going to transfer to grains once they are ready.

Thanks for feedback



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OfflineWorkmanV
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Re: First agar mission [Re: straightslayer]
    #24830224 - 12/06/17 05:45 PM (5 days, 14 hours ago)

The contaminate overgrown dish can be used but there is some risk. Don't use the area where the contaminate was originally found. It would be better to do a transfer into a new plate just to be safe.

You should use them before they reach the edges if possible. The edges are the areas most likely to get contaminated from outside spores. If you let the mycelium grow to the edge and a mold spore gets in and lands on the mycelium, it could just sit there waiting until you transfer into grains to give you trouble.


--------------------
Research funded by the patrons of
The Spore Works
Exotic Spore Supply
Reinvesting 25% of Sales Towards Basic Research and Species Identification :amanitajar:


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Offlinestraightslayer
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Re: First agar mission [Re: Workman]
    #24831318 - 12/07/17 08:05 AM (5 days, 10 minutes ago)

Sweet thank you. They have just started touching edges so I better get to it tomorrow


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Amazon Shop: Agar

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