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OfflineCongame
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Registered: 02/11/20
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No growth on agar dishes (Update: great progress)
    #26480261 - 02/11/20 02:04 PM (3 years, 11 months ago)

Hi everybody,

All new to growing mushroom but I believe I have done a decent amount of research and tried to follow a specific TEK to get it right, but I'm no sure if I'm impatient or what else might me wrong :smile:

So I'm following GordoTEK' 2020 TEK as found on youtube and I have inoculated 8 petridishes with B+ and 8 with GT, I have used a premixed maltextract agar solution that I have bought from a trusted company.
Following Gordo's TEK for autoklave the agar mix first before use, but this actually said on the bottle not to be boiled and since I bought the specifically for mushroom growing i suspect that it is already sterile so I did not autoklave it.
I slowly heated it up till it was all liquid and poured it into the petridishes and let it cool of, of course all done in an as sterile environment as possible.

I did not have my infrared thermometer at hand since I loaned it out to a friend but I waited till dishes wasn't warm too the touch before inoculating the dishes with a drop of spores from sporesyringe.

Then I put dishes in ziplock bags in put them in my incubator set a 82F or 27,5C.

9 days has now past without any signs of growth to both the GT's and B+ dishes, so my question is should i to hold my horses and wait it out or is there a big chance I did something wrong?

The sporesyringes is from one of the biggest european dealers so I have decent amount of trust in that part working but of course that could be the issue as well but highly unlikely with to different strains...
My other two concerns is that the agar might have been to hot after all.. Also I did not shake the sporesyringes before use?!

Hope someone has an idea about what might could have gone wrong here?! :smile:

Thanks!


Edited by Congame (03/11/20 06:25 AM)


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Offlinefeldman114
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Re: No growth on agar dishes after 9 days in incubator [Re: Congame] * 1
    #26480266 - 02/11/20 02:05 PM (3 years, 11 months ago)

Try taking them out of the incubator (no one uses them btw)


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OfflineCongame
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Re: No growth on agar dishes after 9 days in incubator [Re: feldman114]
    #26480277 - 02/11/20 02:11 PM (3 years, 11 months ago)

So you mean it could be too hot?
There is of course not light at all in it but that shouldn't make any difference right?


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Offlinefeldman114
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Re: No growth on agar dishes after 9 days in incubator [Re: Congame]
    #26480290 - 02/11/20 02:15 PM (3 years, 11 months ago)

Light is beneficial at every stage of growth.
The temp we aim for is 72F.

Incubators were deemed useless like 15 years ago. I know you prolly found the tek here, but you generally wanna make sure you follow ones that came out less than 5 years ago.
Those plates might still germinate. I’ve had some take over 2 weeks, in optimum temperatures.


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OfflineCongame
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Re: No growth on agar dishes after 9 days in incubator [Re: feldman114]
    #26480337 - 02/11/20 02:27 PM (3 years, 11 months ago)

Okay, I have pulled the dishes out in to room temp (71F) and fully exposed to light now, say that nothing happens now would it be possible to inoculate the dishes again on the same agar or would the risk of contamination be greater?


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Offlinefeldman114
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Re: No growth on agar dishes after 9 days in incubator [Re: Congame]
    #26480369 - 02/11/20 02:41 PM (3 years, 11 months ago)

Yeah that should work.
Did you drop on top of the agar or use a swab?

Qtips always give me better germination rates.


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OfflineCongame
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Re: No growth on agar dishes after 9 days in incubator [Re: feldman114]
    #26480389 - 02/11/20 02:51 PM (3 years, 11 months ago)

Drop on top


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Offlinefeldman114
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Re: No growth on agar dishes after 9 days in incubator [Re: Congame]
    #26480419 - 02/11/20 03:03 PM (3 years, 11 months ago)

Okay so try this:
Wrap 5 q-tips (individually) in foil and PC for 30ish min.
In the SAB, flame the syringe needle, then unwrap a q-tip and squeeze 2-3 drops of spore solution on the cotton swab.
Use swab to streak a zig-zag pattern on your agar.

No need to push hard or leave wet spots behind - you won’t see the spores but they’re there.


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OfflineCongame
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Re: No growth on agar dishes after 9 days in incubator [Re: Congame]
    #26489554 - 02/17/20 07:25 AM (3 years, 11 months ago)

So finally something started growing!
Sorry for the shitty picture but I couldn't get my camera to focus trough the ziplock bags where I have my petri dishes.



Out of the 14 dishes I have I see growth in one of the B+ once now, it's all white and it's kind of furry at the very edge of the ring... I dont se much growth from the center of the ring and out only a slight blur to the agar, hard to tell in the photo...

So my question is, is it mycelium? :smile:


Edited by Congame (02/17/20 08:52 AM)


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OfflineRaccoon
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Re: No growth on agar dishes after 9 days in incubator [Re: Congame]
    #26489675 - 02/17/20 09:21 AM (3 years, 11 months ago)

It's mycelium, but it does not look like cube mycelium... Shake the syringe and don't use an incubator next time. And it sounds like making your own agar from scratch wouldn't be that much more difficult and cheaper than buying it pre made. Pouring the plates is the pain in the ass part and you still had to do that. That's if you have a pressure cooker, though.


--------------------
First Grow


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OfflineCongame
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Re: No growth on agar dishes after 9 days in incubator [Re: Raccoon]
    #26490093 - 02/17/20 02:08 PM (3 years, 11 months ago)

So this couldn’t be B+ mycelium by any chance?
I’m pretty confident with the vendor of the spore syringes for it to actually being B+ spores, so would this be some foreign spores?


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OfflineCongame
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Re: No growth on agar dishes after 9 days in incubator [Re: Congame]
    #26490350 - 02/17/20 03:57 PM (3 years, 11 months ago)

Took some better pictures:
So no chance this could be B+ mycelium?





Edited by Congame (02/17/20 03:58 PM)


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InvisiblemushboyMDiscord
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Re: No growth on agar dishes after 9 days in incubator [Re: Congame]
    #26490364 - 02/17/20 04:02 PM (3 years, 11 months ago)

I dont think so
:horrified:


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OfflineSockadin
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Re: No growth on agar dishes after 9 days in incubator [Re: mushboy]
    #26490602 - 02/17/20 06:11 PM (3 years, 11 months ago)

Looks like a baby crowning.


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OfflineCongame
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Re: No growth on agar dishes after 9 days in incubator [Re: Sockadin]
    #26491087 - 02/18/20 03:07 AM (3 years, 11 months ago)

And by that you mean? :grin:


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OfflineSockadin
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Re: No growth on agar dishes after 9 days in incubator [Re: Congame] * 1
    #26491143 - 02/18/20 04:42 AM (3 years, 11 months ago)

Loose the incubator and try again with new plates. The above post was referring to giving birth.


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OfflineRaccoon
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Re: No growth on agar dishes after 9 days in incubator [Re: Sockadin]
    #26491466 - 02/18/20 09:50 AM (3 years, 11 months ago)

Even from the best vendor, spore syringes are not perfect. They don't always germinate and they sometimes have contaminates. Because only one plate contaminated, the syringe is probably not contaminated. This contam was probably from a mistake on your part. 7/8 plates not being contaminated means your sterile technique isn't too bad though.

Try again with the same syringe and I hope something happens. I have problems germinating on agar too. Only 4-5 colonies on 20 or so plates. If nothing happens this time around, call the vendor. Good luck.


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First Grow


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OfflineCongame
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Re: No growth on agar dishes after 9 days in incubator [Re: Raccoon]
    #26494420 - 02/20/20 04:54 AM (3 years, 11 months ago)

Thank you guys!

I will try and make some new plates and let then sit at normal room temp. and exposed to light.
When I eventually see mycelium growth I'm planing on picking the best dishes and cut small pieces for agar/mycelium and transfer them to new fresh dishes, from researching around the forum that seems like the way to do it?! I guess I will continued growth pretty fast when the mycelium is already going, right?


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OfflineCongame
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Re: No growth on agar dishes after 9 days in incubator [Re: Congame]
    #26528571 - 03/11/20 06:32 AM (3 years, 10 months ago)

Sooo things are progressing now!
I poured 10 new plates and inoculated them and after 4 days I saw some nice growth I have since made a transfer to some new plates, also I re-inoculated some of my first plates where I didn't see any growth 50% of them showed growth as well and I also did transfers from them...

Now I'm seeing som nice growth on the transfers as well so my question is:
Should I do transfers/sectoring until I see a mono culture or would I be just fine with multi spore, is there a significantly advantage to strive for the mono?
Ps: I'll post some pics of the plates later today!


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