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OfflinePMBastian
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No germination, spores to agar
    #26602904 - 04/15/20 02:42 PM (10 months, 13 days ago)

Just started agar again after doing a couple grows straight mss to grain. My transfers from clones are working fine, but I have trouble getting an ms syringe of GT to germinate.

Thing is, I know the syringe is good, because I've used it for several grain bags that did/is doing ok. But it won't seem to germinate on agar! Now I have 3 plates inoced with the same syringe, one from mid February, one from late Feb and one from a week ago. Nothing has happened on either. No contamination or myc.

Any thoughts? Using oat water diluted 50:50 with tap water, and approx 2% agar. Petris poured in SAB, none of the non-inoced plates shows sign of contam.


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InvisibleRoger Clemency
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Re: No germination, spores to agar [Re: PMBastian]
    #26602950 - 04/15/20 03:05 PM (10 months, 13 days ago)

Perhaps the oat water isn’t giving them the sugars they want, or enough of them? I’ve always used LME and 3-4 day germination on prints. Have you used this recipe for germinating before?


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Offlinepoisoned
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Re: No germination, spores to agar [Re: Roger Clemency]
    #26602952 - 04/15/20 03:06 PM (10 months, 13 days ago)

Yeah, might be better to just go with MEA or potato flakes.


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OfflinePMBastian
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Re: No germination, spores to agar [Re: Roger Clemency]
    #26602961 - 04/15/20 03:11 PM (10 months, 13 days ago)

Could the nutrient really work for cloning without working for germinating? According to a couple of threads I read after searching the forums, whatever medium has nutrients and water should work.

Considering trying another inoc method, scraping dry spores or swabbing them. But I kinda wanna get a culture from that specific syringe!


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Invisiblewoofwoof
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Re: No germination, spores to agar [Re: PMBastian]
    #26602969 - 04/15/20 03:15 PM (10 months, 13 days ago)

You can sterilize some swabs then instead of dropping spore syringe solution to agar, put a drop on the swab then streak your plates.

Also, is your agar hard? haha, softer agar germinates better


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Offlinepoisoned
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Re: No germination, spores to agar [Re: woofwoof]
    #26602977 - 04/15/20 03:18 PM (10 months, 13 days ago)

It's just that there are many possible variables with grain. If you used a well measured out recipe, you could rule that out. Although, I'd imagine cube spores germinating even in water only agar.


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OfflinePMBastian
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Re: No germination, spores to agar [Re: woofwoof]
    #26603018 - 04/15/20 03:32 PM (10 months, 13 days ago)

Quote:

poisoned said:
It's just that there are many possible variables with grain. If you used a well measured out recipe, you could rule that out. Although, I'd imagine cube spores germinating even in water only agar.


yes, I've even had spore syringes germinate.

The recipe is working well for transfers from clone, so in that respect it's well measured out i guess? Kinda winged it based on many GW agar recipes I read on here.

Quote:

woofwoof said:
You can sterilize some swabs then instead of dropping spore syringe solution to agar, put a drop on the swab then streak your plates.

Also, is your agar hard? haha, softer agar germinates better




Do you think swabs would work better? If so, why? I did streak one of the plates with a flamed scalpel, looks just as barren as the two that only had spore water dropped on them...

Agar is pretty soft on the oldest two plates and a tad harder on the newest one.


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Invisiblewoofwoof
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Re: No germination, spores to agar [Re: PMBastian]
    #26603033 - 04/15/20 03:38 PM (10 months, 13 days ago)

I used to use only spore syringe solution because I feared swabbing with agar.
Once I did it, I will never use a syringe straight to agar again. For me, the spore water itself would roll around the plate and it was ugly dirty and had higher contams.

Since swabbing, I have not had any contams. I think the spore water creates nasties


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OfflinePMBastian
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Re: No germination, spores to agar [Re: woofwoof]
    #26603044 - 04/15/20 03:40 PM (10 months, 13 days ago)

That makes sense. Def gonna try it. Just wondering wth is up with this syringe that won't grow either mushrooms or contaminants.


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Invisiblewoofwoof
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Re: No germination, spores to agar [Re: PMBastian]
    #26603053 - 04/15/20 03:43 PM (10 months, 13 days ago)

Quote:

PMBastian said:
That makes sense. Def gonna try it. Just wondering wth is up with this syringe that won't grow either mushrooms or contaminants.



You said the clones are doing good, did you make the agar in same batch?
It could either be the syringe or the PC cycle.
Did you vent the PC?

I use BRF agar.
9g BRF
6g agar (telephone brand)
400ml water


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OfflinePMBastian
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Re: No germination, spores to agar [Re: woofwoof]
    #26603062 - 04/15/20 03:50 PM (10 months, 13 days ago)

Two batches of agar, one soft and one hard. Clones doing good on both. Spores germinating on neither.
I PCd for 30 min and vented the pc, to get the agar out quite hot for pouring.

Hard to compare your nutrient ratio to mine, as I used grain water. I did use more agar than you tho, 2% first batch and 2.5% second. Telephone brand agar. But as the clones and non-inoced plates are doing well, it doesn't make sense to me that something is wrong with sterilisation/recipe.


Edited by PMBastian (04/15/20 03:51 PM)


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InvisibleInfiniteDreams
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Re: No germination, spores to agar [Re: woofwoof]
    #26603063 - 04/15/20 03:51 PM (10 months, 13 days ago)

I've germinated spores on oat water agar.  Exact same timeframe as MEA (4-5 days) to see visible mycelium.


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OfflinePMBastian
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Re: No germination, spores to agar [Re: InfiniteDreams]
    #26603067 - 04/15/20 03:53 PM (10 months, 13 days ago)

Quote:

InfiniteDreams said:
I've germinated spores on oat water agar.  Exact same timeframe as MEA (4-5 days) to see visible mycelium.




Bah, I guess I'll just have to give up on this syringe then. Even though it produces when put straight to grain. Guess I'll try swabbing with a new sporeprint when I have time!


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InvisibleInfiniteDreams
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Re: No germination, spores to agar [Re: PMBastian]
    #26603075 - 04/15/20 03:56 PM (10 months, 13 days ago)

Don't know what to tell you.  Personally I wouldn't toss the syringe if you know it is good. 

Did you shake it up really well before applying the drop to the agar?

Also FWIW, my oat water came from a 45 minute boil so it is probably nutrient rich.  For MEA I tend towards a 1.5% nutrient.


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OfflinePMBastian
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Re: No germination, spores to agar [Re: InfiniteDreams]
    #26603089 - 04/15/20 04:02 PM (10 months, 13 days ago)

Yup, shook the hell out of it.

I meant giving up germinating that syringe on agar. Don't have enough cultures on agar to keep the wheels turning, so I still have to noc up a bag with mss every once in a while.

My oat water is from a 45-50 min boiling water soak. So probably less nutritious than yours (bc it didn't boil continously) and I diluted it with equal parts water, after reading that less nutes are better than too much and that several people did this with grain water.


Edited by PMBastian (04/16/20 02:32 PM)


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