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OfflineFrrrunkis
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I'm a teapot
Registered: 06/14/20
Posts: 110
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Re: Syringe to agar - need advice after 1 week [Re: Kyngwhatt]
    #26878747 - 08/13/20 02:44 PM (3 years, 5 months ago)

Behold, round two. I've got many more of these now, but I'm only showing the interesting ones. I've started labelling them slightly better too. Excuse the condensation, I'm still not good at pouring at the right temperature. :/

So, remember 7 and 9 from the first batch? Here they are today!

Adam and his 2 kids:

I'm pretty sure they're not cubes. Cobweb mold?

Eve and her 2 kids (please appreciate the unintended vagina):

Same story as with Adam. All of these plates grew super fast too. I'm leaving them all to turn into whatever.

So, these are fresh from that same syringe as the first round, but I gave it a good shake this time. I made 8 of these, of which 3 are looking promising:

I think I'll try and make transfers (grain spawn material) from all 3 of these, at least 5 transfers each. Is it ok to transfer now (in 2-3 days), or should I wait longer?

I also got this piece of modern art. Any takers?


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Offlineverum subsequentis
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Re: Syringe to agar - need advice after 1 week [Re: Frrrunkis]
    #26878764 - 08/13/20 02:53 PM (3 years, 5 months ago)

Contam bonanza. I see nothing worth saving in these pics. I believe you have a contaminated syringe and need to work on your sterile tek.


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OfflineFrrrunkis
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Re: Syringe to agar - need advice after 1 week [Re: verum subsequentis]
    #26878789 - 08/13/20 03:15 PM (3 years, 5 months ago)

Ouch. Aren't all 3 white growths in the 3rd row cube mycelium? It's hard to take a photo through the condensation, but it looks white and fluffy, plus they all formed around black dots (spore clumps?) from the syringe.

As for the syringe and sterile tek, I'm sure you're right. I want to try and salvage what I can though.


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Literally nothing matters. This is fine.


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OfflineFrrrunkis
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Re: Syringe to agar - need advice after 1 week [Re: Frrrunkis]
    #26883505 - 08/16/20 01:48 PM (3 years, 5 months ago)

Ok, I'd really appreciate another look. I hope these photos provide more detail.

02/08//3


02/08//5


02/08//4


I'm not sure which of the spots/discolorations on agar are normal (condensation spots/streaks, agar or malt extract clumps) and which are contaminants.

Looking closely, there seem to be sections where I should be able to cut just the white mycelium.



Edited by Frrrunkis (08/16/20 01:56 PM)


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Offlinedirtybirdx
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Re: Syringe to agar - need advice after 1 week [Re: Frrrunkis]
    #26883535 - 08/16/20 02:06 PM (3 years, 5 months ago)

Yea man I’d be afraid to continue with any of those...

;(

There is a bit of healthy mycelium surrounded in a few places but it wouldn’t be worth it in my opinion.


Did you make/use a still air box?  Even the first 50 plates I made never grew anything that funky.


You might be bummed about shroomies being pushed back, but this is quality learning for you.  When you see the healthy white rhizo mycelium, you’ll feel much better.

A lot of molds are white.  Cobweb mold which is very common shows up a lot with us and I’ve probably even thrown away good cultures being unsure and not wanting to take the risk.

That’s why rhizomorphic growth is so coveted — it’s so easy to see whether it’s contaminated or not.  Gives you more of an assured feeling.

Toss them, make your still air box, pour some more plates, and give it another go!  👍


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Offlinedirtybirdx
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Re: Syringe to agar - need advice after 1 week [Re: dirtybirdx] * 1
    #26883546 - 08/16/20 02:18 PM (3 years, 5 months ago)

Also, you said you weren’t sure what you were seeing, so

- you have bacterial contamination around the entire outer edge of every plate.  It’s the whitish yellowish film near the sides.

That’s not necessarily a plate-killer, but the fact that it’s present in all three sounds like sterile technique, or potentially unsterilized agar.

- most of the white you see in the first two plates are mold.

- the ring on the third plate, and most of that growth, is mold.

- yellow spot is bacteria

- the only real mycelium I’m confident about is on the second plate, you see two small growths resembling hands - that’s what you want the whole growth to be.  And with none of the other stuff.

Hope this helps


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OfflineFrrrunkis
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Re: Syringe to agar - need advice after 1 week [Re: dirtybirdx]
    #26883572 - 08/16/20 02:35 PM (3 years, 5 months ago)

Thanks for your input!

Yeah I use a SAB, as per Bod's tek. I've only worked in it 3 times so far (this was my 2nd agar pour/use), so I'm clumsy and my technique sucks, but I think I'm as prepared as possible. Gloves, iso, flame, loop, scalpel, the whole thing. I sterilize my agar in a PC for 30 mins. Glass petris are oven-sterilized at 250C for 30 mins. I've now bought sterile plastic petris to simplify the process.

Maybe this syringe is really just bad and I should get another one, or a spore print.

I guess I'll still try to transfer from these plates just to see what happens. I'll give my syringe another fresh go too, just to be sure. Maybe I'll get lucky. If not - at least I'm learning.

The "whitish yellowish film" on the outer edge is only there on 02/08//4. On the oher two that's actually the parafilm wrap (it reaches around the edge onto the top and bottom a bit, and it's semi-transparent, so it looks similar under the light). So hopefully at least my agar is clean.


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OfflineFrrrunkis
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Re: Syringe to agar - need advice after 1 week [Re: Frrrunkis] * 1
    #26969138 - 10/04/20 01:49 PM (3 years, 3 months ago)

I just wanted to thank you guys for the advice I've received here! I gave these plates a shot, did 2 more rounds of transfers and I've now got 7 oat jars going strong. Today I was going through all my plates to see what's staying and what's not, and - to my surprise - some of them are pinning!



I'm now much more confident that I can actually make this work.

Thanks again!


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Offlineverum subsequentis
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Re: Syringe to agar - need advice after 1 week [Re: Frrrunkis]
    #26969190 - 10/04/20 02:15 PM (3 years, 3 months ago)

congrats


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OfflineAtmozFear
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Re: Syringe to agar - need advice after 1 week [Re: Frrrunkis]
    #26969205 - 10/04/20 02:20 PM (3 years, 3 months ago)

I still have yet to see pins from an agar plate


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Offlineverum subsequentis
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Re: Syringe to agar - need advice after 1 week [Re: AtmozFear]
    #26969217 - 10/04/20 02:26 PM (3 years, 3 months ago)

It'll happen.


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Offlinehazyhorse
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Registered: 03/19/19
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Re: Syringe to agar - need advice after 1 week [Re: verum subsequentis]
    #26969994 - 10/05/20 12:18 AM (3 years, 3 months ago)

just forget abt em long enough & they pop up lol when i get lazy it starts to happen

congrats on the agar


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