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OfflineRotnpins
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Re: Let's talk about Water Agar!! ๐Ÿ’ง [Re: Rotnpins]
    #27998799 - 10/14/22 02:35 PM (1 year, 3 months ago)

:offthehook:


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InvisibleBaba Yaga
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Re: Let's talk about Water Agar!! ๐Ÿ’ง [Re: Rotnpins] * 2
    #27998823 - 10/14/22 02:45 PM (1 year, 3 months ago)

The pan pin is showing good recovery in the mycelium department while bacterial goop doesn't seem to thrive.




That is already a win in my book. Working better than tee agar so far.


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OfflineRotnpins
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Re: Let's talk about Water Agar!! ๐Ÿ’ง [Re: Rotnpins]
    #27998826 - 10/14/22 02:47 PM (1 year, 3 months ago)

:bearbreakdance:


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InvisibleBaba Yaga
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Re: Let's talk about Water Agar!! ๐Ÿ’ง [Re: Rotnpins] * 1
    #28001887 - 10/16/22 02:51 PM (1 year, 3 months ago)

5 days after the drop



Easy peasy, thanks for bringing this to my attention mate.


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InvisibleEgon_Spengler
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Re: Let's talk about Water Agar!! ๐Ÿ’ง [Re: Baba Yaga] * 2
    #28002401 - 10/16/22 09:03 PM (1 year, 3 months ago)



Oh yeah, this is that same RW, after being back on nutes for 5 days. Itโ€™ll go to grain once fully grown out. Probably next weekend.

THANKS AGAIN!


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InvisibleSoutherner
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Re: Let's talk about Water Agar!! ๐Ÿ’ง [Re: Egon_Spengler] * 2
    #28014382 - 10/24/22 04:14 PM (1 year, 3 months ago)

First off, thanks for the write-up Rotn. Iโ€™ve read a few people talking about water agar before but I think this thread has really brought more attention to the technique.

Wanted to show yโ€™all this and see if you had experienced it before.



Those cloudy areas around the center and left developed after I had taken the wedges you see cut out. Wondering if itโ€™s a delayed bacterial bloom. The black pearl oyster I put to water agar did not develop this.

Another thing Iโ€™d like to mention is as I was wrapping the plates I poured the bottom one slipped off and popped open. Said fuck it and opened it up and waved it around a bit before sealing. NOTHING grew on it. So although this is a nice tool in the toolbox still be careful. You may still be transferring nasties if you are sloppy.


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OfflineRotnpins
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Re: Let's talk about Water Agar!! ๐Ÿ’ง [Re: Southerner]
    #28014445 - 10/24/22 04:43 PM (1 year, 3 months ago)

Glad you enjoyed the write up :super: it's really awesome to see more people giving this a try.

I just wanted to confirm that I'm on the same page before I answer your question :lol: are you referring to the areas I marked in red?



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InvisibleSoutherner
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Re: Let's talk about Water Agar!! ๐Ÿ’ง [Re: Rotnpins]
    #28014466 - 10/24/22 04:53 PM (1 year, 3 months ago)

:lol: I could have done some better explaining on that but yes the areas you marked are what Iโ€™m wondering about.

Yeah Iโ€™m really eager to try cloning with water agar as bacteria is usually the biggest issue. Hopefully will find some wild fruits after this nice rain to try with.


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OfflineRotnpins
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Re: Let's talk about Water Agar!! ๐Ÿ’ง [Re: Rotnpins]
    #28014511 - 10/24/22 05:11 PM (1 year, 3 months ago)

I've seen this type of growth on a bunch of water agar plates.. I don't think it has to do with bacteria.. I believe that it's just stronger mycelium growth than the rest of the plate..

In my "nutrient sample" experiment I ran a couple pages back, I put a small piece of nutrient rich agar on a WA plate to see how the mycelium would react when it reached the sample.. it ended up making a ring around most of the plate with that type of growth... the nutrients gave the mycelium a temporary food source resulting in a small ring of stronger growth..

I've also seen the same growth on plates with only WA and no nutrients.. both, while the plate was growing out, and later on when the plate was grown out most/all of the way...

when it shows up at the end, I think it's the mycelium reacting in a similar way to some of the stranger growth that tends to show up on overgrown nutrient plates. And when it shows up while the plate is growing out, I usually take it as a good indicator of healthy mycelium (as long as it's far enough from the original transfer).. when it shows up early on a plate, and I'm still concerned that it's too close to the original transfer, I usually assume that it's either from the nutrients in the transfer wedge or the mycelium trying to outrun the bacteria, but I haven't done any tests yet to confirm..

I've tested the theory that it is strong growth (when far enough from the transfer) and not due to bacteria by taking transfers from those areas.. so far all of those transfers have been a success and resulted in clean plates.

I'm still learning and testing different theories about growth patterns on WA plates, but it's some food for thought.


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InvisibleSoutherner
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Re: Let's talk about Water Agar!! ๐Ÿ’ง [Re: Rotnpins]
    #28014574 - 10/24/22 05:31 PM (1 year, 3 months ago)

Thanks for the explanation. Iโ€™m not entirely sure if the culture is contaminated but I feel like it might be. With the intertwined bacteria or mold I find it hard to tell on agar. Iโ€™m sure more experienced growers have a better eye for it.

I wonder if testing a bacterial enmeshed culture in LC would be better. Might better show itself as cloudiness as opposed to hiding in the myc.


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OfflineRotnpins
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Re: Let's talk about Water Agar!! ๐Ÿ’ง [Re: Southerner] * 2
    #28014650 - 10/24/22 06:13 PM (1 year, 3 months ago)

Do you have any pictures of the new plates?

Edit: fixing a typo because rusty is nit-picking
:godfuckingdammit:


Edited by Rotnpins (10/24/22 06:49 PM)


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OfflineRusty2096
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Re: Let's talk about Water Agar!! ๐Ÿ’ง [Re: Rotnpins]
    #28014719 - 10/24/22 06:45 PM (1 year, 3 months ago)

Quote:

Rotnpins said:
Do you have any pictures if the new plates?




Dyslexia 1, Rotn 0


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InvisibleSoutherner
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Re: Let's talk about Water Agar!! ๐Ÿ’ง [Re: Rotnpins] * 2
    #28016051 - 10/25/22 02:22 PM (1 year, 3 months ago)

Fuck it, I needed to pour some agar today anyways so did a couple LCs as well. I donโ€™t know what this will prove if it even shows anything but figured it couldnโ€™t hurt.



Edit - Yikes: Better Late Than Never Update 1/19



Both the light and thickened areas grew out the same in LC in line with what Rotn suggested.

250ml distilled water
.25g LME (.1%)
PCd 30 min at 15+ PSI



The larger wedge next to where A is written on the dish is what I actually used for the LC. One from a cloudy section one not. Wish I had identical jars as the other one has less clarity but weโ€™ll see what happens.

Quote:

Rotnpins said:
Do you have any pictures of the new plates?

Edit: fixing a typo because rusty is nit-picking
:godfuckingdammit:




I do but itโ€™s a gourmet so not sure how many folks on this side have experience with it. I asked in the gourmet section and got conflicting opinions.



Pretty aerial closer to the middle. Not sure if thatโ€™s a sign of contamination, aggressiveness or possibly how it looks on 2% LME.


Edited by Southerner (01/19/23 03:37 PM)


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OfflineRotnpins
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Re: Let's talk about Water Agar!! ๐Ÿ’ง [Re: Rotnpins] * 1
    #28018032 - 10/26/22 05:36 PM (1 year, 3 months ago)

Here's a new one for you guys...

Jack Frost knotting up on water agar plate


@southerner... note the growth that looks similar to what we were talking about.. JF throws out really strong rhizo, so it's very easy to see all the strong growth.


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OfflineRotnpins
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Re: Let's talk about Water Agar!! ๐Ÿ’ง [Re: Rotnpins]
    #28018052 - 10/26/22 05:45 PM (1 year, 3 months ago)

Extremely bacterial Tidal Wave clone that I sent to WA for clean up...
It's showing visible bacteria on the outer edge of the plate, like I said, the sample was really bacterial :lol:



Knots showing up on the clean side of the plate


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InvisibleSoutherner
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Re: Let's talk about Water Agar!! ๐Ÿ’ง [Re: Rotnpins] * 1
    #28018059 - 10/26/22 05:47 PM (1 year, 3 months ago)

Holy shit, thatโ€™s awesome. Fungi are some badass life forms.

You talking about the brown area for the bacterial spot?


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OfflineRotnpins
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Re: Let's talk about Water Agar!! ๐Ÿ’ง [Re: Southerner] * 1
    #28018078 - 10/26/22 05:54 PM (1 year, 3 months ago)

Quote:

Southerner said:

You talking about the brown area for the bacterial spot?




Yupp.. it's the only thing that makes sense :shrug: this is the first time I've had a plate that didn't get 100% clean by the time it got to the outer edge..

very cool that it seems to be easily identifiable when it's not clean :awesomenod:


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OfflineRotnpins
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Re: Let's talk about Water Agar!! ๐Ÿ’ง [Re: Rotnpins] * 5
    #28018148 - 10/26/22 06:33 PM (1 year, 3 months ago)

One last photo drop from today's agar session...

The original Enigma plate I was sent was bacterial and had visible metabolites.. I ended up cleaning it up with WA and sent a transfer back to nutrient agar.. that plate produced the 2 successful bags of Enigma that I ran (they were a little bacterial, but it was from a grain prep failure. The nutrient plate was clean).. so I'm fairly confident that all but the center of this plate is clean..

I decided to send some of my old WA plates to grain to see how things went..

Here is the Enigma plate:


This is how I cut the plate.. the goal is to leave the center (and hopefully the contamination) behind and use the rest of the plate:


Here is a shot of the jar I inoculated:


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OfflineCJD
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Re: Let's talk about Water Agar!! ๐Ÿ’ง [Re: Rotnpins] * 1
    #28018166 - 10/26/22 06:50 PM (1 year, 3 months ago)

That looks like a fun experiment. I have to admit it's hard to see the mycelium in the photos but once you make it out you can see it.  I just poured a bunch of WA condiment cups.  I plan to use them with 5 new varieties I just germ plated. Can't wait to see some life in those.


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OfflinePanMax
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Re: Let's talk about Water Agar!! ๐Ÿ’ง [Re: Baba Yaga]
    #28019980 - 10/27/22 06:27 PM (1 year, 2 months ago)

Quote:

Baba Yaga said:
The pan pin is showing good recovery in the mycelium department while bacterial goop doesn't seem to thrive.




That is already a win in my book. Working better than tee agar so far.




Does dropping a pin on to agar work for cloning??


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