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Offlinejpchatx
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Big white dot on agar
    #27454193 - 09/02/21 09:44 PM (3 years, 4 months ago)

I’m assuming the big white dot is a contam, but this is my first time doing agar from a spore syringe—no transfers yet. The smaller dot seems like it could be healthy myc, any thoughts? I mostly want to make sure I’m not transferring Contams.




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OfflineInsultingLizard
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Re: Big white dot on agar [Re: jpchatx]
    #27454204 - 09/02/21 09:53 PM (3 years, 4 months ago)

They both look like mold to me. Just-germinated spores are always flat, not with volume. Give it some time, but I'm pretty sure they'll turn green or black in the next 48 hours.

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OfflineVibe_Enthusiast
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Re: Big white dot on agar [Re: InsultingLizard]
    #27454217 - 09/02/21 10:06 PM (3 years, 4 months ago)

Think I'm going to have go with :whathesaid: looks like they're both rising. Almost like a cloud.

You'll know for sure within a few days. If it turns out to be healthy myc.. maybe you can transfer before it meets with the contam.

Although, I'm hoping when you knock plates up you do more than one. I usually do a batch of 20 agar dishes and drop spores to about 6 of them. Always good to have backups.. and always good to take transfers of fast healthy growing mycelium!

Good luck!


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:greyalien:



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Offlinejpchatx
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Re: Big white dot on agar [Re: Vibe_Enthusiast]
    #27454221 - 09/02/21 10:13 PM (3 years, 4 months ago)

Ah perfect, okay. I did see the volume on it and wasn’t sure but that’s a good tip. I only did 3 plates, and looks like 2 have those big fast growing spots. The third I don’t see any growth yet, so I’m hoping that’s the good one. But if not I have 17 left :cool:

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OfflineVibe_Enthusiast
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Re: Big white dot on agar [Re: jpchatx]
    #27454224 - 09/02/21 10:15 PM (3 years, 4 months ago)

Are you using a SAB when dropping spores?


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:greyalien:



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OfflineInsultingLizard
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Re: Big white dot on agar [Re: jpchatx]
    #27454232 - 09/02/21 10:21 PM (3 years, 4 months ago)

I just noticed the dates. You inoculated these on the 26th? So that took a week to grow? Mmh... I'm a bit on the fence on this one, so just watch them.

By the way, are you streaking the plates? When working with a doubtful print or solution you want to do that to spread out whatever is on it over the plate, so you can separate a useful colony from contamination.

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OfflineFestivusmiracle
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Re: Big white dot on agar [Re: InsultingLizard]
    #27454303 - 09/02/21 11:54 PM (3 years, 4 months ago)

I wouldn’t give up on these especially if it took a week or so to grow.  Germination plates are ugly and often very tomentose with the hundreds of strains trying to take off.  My germination plates looked similar.  It wasn’t until several transfers that I had something that looked more like the plates people show off here.  If you want more seasoned advice post them on the agar envy thread.  I have hope its mycelium and wouldn’t ditch them yet.  Pics of my plates that I thought were mold but I transferred and had several successful harvests from.


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OfflineFestivusmiracle
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Re: Big white dot on agar [Re: Festivusmiracle]
    #27454306 - 09/02/21 11:57 PM (3 years, 4 months ago)

That is a helpful point.  If you streak the plates and do it hard enough that it marks the agar you can tell if what’s growing came from your streak or is a satellite contam.  I still bet that is mycelium.  You will know for sure in a transfer or 2.

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Offlinejpchatx
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Re: Big white dot on agar [Re: Festivusmiracle]
    #27460591 - 09/08/21 02:49 PM (3 years, 4 months ago)

Ok this is super helpful. I figured there was no harm in keeping to watch how they developed and I feel like 01 is myc with with a contam (that solid round looking guy looks suspicious but the others are kind fluffy), and maybe 02 is un contaminated?? Or maybe I have the start of a healthy mold farm.

I didn’t streak unfortunately but I am doing this in a SAB with the best clean technique I can manage. I have 17 extra plates.

My plan is to do a transfer with these and see what happens. Should I also reinoculate a clean plate or two to be safe? I have 17 to spare and am not too worried about specific strain isolation at this point, this is just my first time doing agar.


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OfflineInsultingLizard
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Re: Big white dot on agar [Re: jpchatx]
    #27460621 - 09/08/21 03:20 PM (3 years, 4 months ago)

The fuzzy colonies on 01 and 02 are definitely mushroom mycelium.

01: What color is that little dot in the middle of the sharp circular colony? I can't quite make it out. Is it green, or is it the color of the agar? Can you get another shot of this dish from a lower angle? I want to see if the colony has volume.

03: I'm not sure I like that colony too much, but if it's only that big after almost two weeks and it's still white it's probably not mold.

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Offlinejpchatx
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Re: Big white dot on agar [Re: InsultingLizard]
    #27460669 - 09/08/21 04:11 PM (3 years, 4 months ago)

03 looks significantly different than the others, it’s compact, uniform and probably half the size. But pure white.

The dot in 01 is definitely greenish. There’s a strong looking segment in 01 at about 4 o’clock (if the writing is level) that looks worth grabbing, but not sure if it’s too late given the development of the green patch. I tried to do an angled shot but my phone refuses to focus correctly, not sure how useful it is.

I’m looking at grabbing the two fingers my actual finger is pointing to in 02 also. Any other tips? I’ll post my results after the transfer.


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OfflineInsultingLizard
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Re: Big white dot on agar [Re: jpchatx]
    #27460717 - 09/08/21 04:59 PM (3 years, 4 months ago)

Yeah, the really sharp one on 01 is definitely mold. It's odd how little it has sporulated. You can transfer from that dish, but just be careful not to jostle it too much.
Also, in the future try to make sure the surface of the agar is in focus.

Don't worry about selecting any specific sectors from those colonies. They're so tomentose because there's hundreds or even thousands of strains in there, so one way or another you're going to get lots of strains no matter where you transfer from.

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Offlinejpchatx
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Re: Big white dot on agar [Re: InsultingLizard]
    #27460974 - 09/08/21 08:26 PM (3 years, 4 months ago)

Great, thanks for the tips. I’m doing the transfer tonight!

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Invisiblec10h12n2o
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Re: Big white dot on agar [Re: jpchatx]
    #27461196 - 09/09/21 02:46 AM (3 years, 4 months ago)

how are you inoculating these plates?

id suggest doing a serial streak like in my sig and in my agar guide, it will give you lots to choose from and is super helpful for separating from contams



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