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OfflineaTyNieee
Registered: 05/01/20
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First time everything. Spores to agar
    #26778405 - 06/24/20 04:46 AM (17 days, 3 hours ago)

Hello everyone! Thanks to MrBovineJoni user of this forum I get some spores.
I used light malt extract 10g, agar-agar 10g and nutritional yest flakes 1g to prepare ~400ml agar. I think using nutritional yest flakes was a  first mistake. They did not melted and after 20min of sterilization process there was sediment in a media bottle. The agar I poured was probably too hot (second mistake) which caused condensation on the petri dishes lids, so I tried to get rid of it by putting a dish with hot water on the top. It helped a bit. After that I transferred spores to agar. After 3 days I end up with this:







which looks like growing mold to me. Am I right? Is the temperature of agar and sediment in the media bottle the only reason of it? Can I 'save' this plates somehow or maybe that's better to throw them away without opening them at all?


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OfflineGastronomicus
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Re: First time everything. Spores to agar [Re: aTyNieee]
    #26778435 - 06/24/20 05:34 AM (17 days, 2 hours ago)

That is mold yes. Toss em since your spores haven't germinated yet. Since you got it so quickly either something is wrong with your sterilization initially, or your technique during inoculation. Do you have other plates that you made that you haven't opened yet? Are they still mold free?

Don't get discouraged. My first plates grew nothing but mold too. Watch some SAB videos and practice your technique.


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Make my Funk the P Funk, I wants to get Funked up


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OfflineaTyNieee
Registered: 05/01/20
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Re: First time everything. Spores to agar [Re: Gastronomicus]
    #26778560 - 06/24/20 07:01 AM (17 days, 1 hour ago)

I've got plates with hawaiian and these ones with keepers creepers. Mold seems to be in most of kc plates, but it looks like spores have started germinated in these plates (looking from the bottom). Hawaiian have not started growing yet at all (there is few plates with mold, but not so manny as in kc plates). The agar I've used was same for both of them. I prepared 2 media bottles and split each of them for kc and haw.


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OfflineGastronomicus
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Re: First time everything. Spores to agar [Re: aTyNieee]
    #26778596 - 06/24/20 07:23 AM (17 days, 1 hour ago)

Ok so if only those plates are contaminated and the rest from the same batch aren't it means that you messed up your sterile technique in the SAB. Probably let your hand cross over the open plate. It takes some getting used to for sure.

But definitely toss those plates you posted ASAP. The green you see are spores and you don't want those getting out in your grow area


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Make my Funk the P Funk, I wants to get Funked up


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OfflineaTyNieee
Registered: 05/01/20
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Re: First time everything. Spores to agar [Re: Gastronomicus]
    #26778618 - 06/24/20 07:34 AM (17 days, 50 minutes ago)

Thank you for your help. I will get rid of these plates.

What about these ones?



I have got also a question about SAB. I've got a 60ltr box (I'm going to but something about 100ltr, because the one I've got is a bit too small) which I have used with 2 small rack in it. Should I use a towel too (below the racks)? Should I soak it in water mixed with soap? What about walls of SAB? I cleaned them with 70% ISO trying to move a paper towel in one direction. Is that enough?


Edited by aTyNieee (06/25/20 01:30 AM)


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OfflineGastronomicus
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Re: First time everything. Spores to agar [Re: aTyNieee]
    #26783946 - 06/26/20 08:30 AM (14 days, 23 hours ago)

Those plates look good. Take some transfers from the cleanest looking sections as soon as you can.

I personally do not use a towel but some people do. I don't think it matters too much.


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Make my Funk the P Funk, I wants to get Funked up


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OfflineaTyNieee
Registered: 05/01/20
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Re: First time everything. Spores to agar [Re: Gastronomicus]
    #26791314 - 06/29/20 03:51 AM (12 days, 4 hours ago)

Is this not a cobweb in these plates? How can I figured it out?

Once again thank you for your help!


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OfflineGan
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Re: First time everything. Spores to agar [Re: aTyNieee]
    #26791474 - 06/29/20 06:49 AM (12 days, 1 hour ago)

Well not sure what your plates look like now. But cobweb isn't as white as cube myc, it grows much more wispy than cube myc, much less organized, and much, much faster. Along with some other tell tale signs.


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OfflineTrichodingus
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Re: First time everything. Spores to agar [Re: aTyNieee]
    #26791670 - 06/29/20 09:12 AM (11 days, 23 hours ago)

Quote:

What about walls of SAB? I cleaned them with 70% ISO trying to move a paper towel in one direction. Is that enough?





Here is what I do:

Turn off house fan, AC, heat etc to reduce air currents
Wash the box inside and out.
Dry completely with paper towel.
Set box up with everything inside.
Cover tools with foil
Mist the inside of box with water
Wait 10 minutes
Success

IME it's more important what your hands do inside the box than how you clean it.


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OfflineaTyNieee
Registered: 05/01/20
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Re: First time everything. Spores to agar [Re: Gan]
    #26792140 - 06/29/20 01:51 PM (11 days, 18 hours ago)

I will take some photos tomorrow morning. I'm a newbee, so I might be wrong. It looks like all of the Keepers Creepers plates are covered with coweb. They started growing really fast. It's been over a week now since I've transferred spores to agar. I have got plates with hawaiian and the ones with KC. And the difference between them is massive.


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OfflineaTyNieee
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Re: First time everything. Spores to agar [Re: aTyNieee] * 1
    #26793753 - 06/30/20 05:31 AM (11 days, 2 hours ago)

Few photos of my plates:


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OfflineMrBovineJoni
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Re: First time everything. Spores to agar [Re: aTyNieee]
    #26794157 - 06/30/20 10:06 AM (10 days, 22 hours ago)

Looking good! In pretty much all of your plates I can see mycelium growing (others more rigorous than others).

That very fluffy Hawaiian looks a bit odd but that does not immediately mean it is bad. If you have some plates to spare for transfers, take a transfer from it and see how it looks like when it recovers.


If this the same KC I sent you (got it from Edmunter myself) - I would expect it to be growing quite  quickly (and also the weather has been very warm lately). But be careful as one of those richer KC plates has some mold on the side so try to transfer without dragging that along.


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OfflineaTyNieee
Registered: 05/01/20
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Re: First time everything. Spores to agar [Re: MrBovineJoni]
    #26794236 - 06/30/20 10:41 AM (10 days, 21 hours ago)

Thank you once again mate!

Everything on these plates is what I received from You (excluding mold on the one)

I noticed that darker area on one plate and I thought it does not look good. I have prepared lots of agar plates using the recipe you have sent me. I will do transfers tomorrow :smile:


Edited by aTyNieee (06/30/20 10:48 AM)


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OfflineaTyNieee
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Re: First time everything. Spores to agar [Re: aTyNieee] * 1
    #26814232 - 07/10/20 04:00 AM (1 day, 4 hours ago)

I've made few transfers a week ago. I would like to ask you what you think about them.

Are these bacterial? I am asking about that clearer (I am not sure how to describe it) area on the agar, which seems to stop mycelium from growing. It is the most visible on the first and the second plate.



Is that cobweb mold growing on the left hand side on this one?


I believe this one looks perfect:



Should I use this one to make more transfers?


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Offlinew00tmycelium
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Re: First time everything. Spores to agar [Re: aTyNieee]
    #26814237 - 07/10/20 04:06 AM (1 day, 4 hours ago)

Are those T1 transfers or what?  The later pictures look amazing but I'm still new to this too.  Also you might be right about the cobweb mold, I'd just cut that section out heavily and keep it going for as long as you intended.


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OfflineaTyNieee
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Re: First time everything. Spores to agar [Re: w00tmycelium]
    #26814257 - 07/10/20 04:18 AM (1 day, 4 hours ago)

First transfer - spores to agar. You can find pictures of T1 at the beginning of this topic (clear/yellow agar plates). That one is T2 (green agar).

I am not sure about this section with cobweb. It might be safer to just do another transfer from that plate. I'm a newbee too :wink:


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OfflineMrBovineJoni
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Re: First time everything. Spores to agar [Re: w00tmycelium] * 1
    #26814266 - 07/10/20 04:24 AM (1 day, 4 hours ago)

Good job man. Half of those plates look really good (4,5,7 from the first row) and the rest look quite decent too. And the last picture  indeed looks perfect.

That plate with the contamination on the side- don't think it's cobweb (it's normally a lost more wispy looking) - however, the mycelium looks healthy so you can make a few transfers on a new plate.


I don't think you need to transfer the perfect plate more times - I would let it grow out a little bit more and use agar slices to inoculate grains already. I probably would also do that with 5&7 from the first row of pictures if they still look good after a few days (after the mycelium covers > 2/3 of the plate area, looks healthy and does not touch walls(not strictly required but that usually is safer)).

Well done!


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OfflineaTyNieee
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Re: First time everything. Spores to agar [Re: MrBovineJoni] * 1
    #26814277 - 07/10/20 04:29 AM (1 day, 3 hours ago)

Thank you so much mate! I would not be able to achieve this without your help  :bow2:  :bow2:  :bow2:


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Offlinew00tmycelium
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Re: First time everything. Spores to agar [Re: aTyNieee]
    #26814281 - 07/10/20 04:34 AM (1 day, 3 hours ago)

You've inspired me even more... I hope to get plates that look close to that soon.  But this is probably a really stupid question, how can I link pictures side by side like you just did?  Last time I pasted BBcode given by the forum upload page it was all vertical.

EDIT: Also someone told me here this week that T0 was spore to agar, and agar to agar was T1.  Not sure which one use yet to describe them..


Edited by w00tmycelium (07/10/20 04:36 AM)


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OfflineLogicaL ChaosM
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Re: First time everything. Spores to agar [Re: aTyNieee]
    #26814326 - 07/10/20 05:13 AM (1 day, 3 hours ago)

Regarding your most recent pics: The first 3 plates look bacterial (the slimy-clearish stuff) but the other plates look nice and clean! Nice work!


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