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Let's talk about Water Agar!! 💧 * 31
    #27900393 - 08/12/22 10:03 AM (1 year, 5 months ago)

When I first started working with agar, I had an abundance of plates that I thought were too contaminated to clean up.. I thought about using antibiotics, but i had read that it could also hinder the mycelium growth and I didn't want to get stuck using it as a crutch.. So I decided to try making some water agar..

What is "water agar"?
Water agar is exactly what it sounds like, water and agar without any added nutrients.. the mycelium grows out very quickly (compared to the contams) looking for food.. this gives it a chance to out grow any contamination that may be riding along.

How to make water agar:
The recipe I use is 7g Agar powder per 500ml H2O

I leave the plates clear and add a different food coloring to each of my nutritional plates so I can easily distinguish between the type of plates.. (for example: H2O Agar=clear, MEA= blue, YMEA= yellow, etc,.)



You would prepare the water agar the same way as your nutritional agar plates.. the method I use is this:

1: add 100ml cold/room temp H20 to media bottle.
2: measure out remaining water (400ml for a 500ml batch) and heat water using preferred method. I personally bring it to a light boil on the stove.
3: while the water is heating, measure out your agar powder (7g per 500ml) and add it to your media bottles. Then give the bottle a little bit of a shake/stir.
4: once it is hot, add the remaining water to the media bottle. Put the lid on and shake it up.
5: prepare your PC with enough water for a 20 minute cycle.. put the water agar media bottle in the pc. If you have any nutritional agar, instruments that need sterilizing, no pour plates, etc,. you can throw them in the PC at the same time.
6: vent steam from PC for 10-15 minutes, add your weight, bring PC up to 15-17psi, reduce heat to low,, set a timer ⏲ and let cook for 20 minutes.
7: when PC cycle is done, turn the heat off and wait for pressure to drop to 0 on its own.
8: take your bottle(s) to your work space and wait for them to cool to pouring temp (I usually pour between 140-120 degrees)

How to use water agar:
Once your plates have cooled, it's time to do some transfers..

I make my transfers the same way I would when transferring to a nutrient plate.. I find an area that seems to have the most organized growth and I take a small transfer to the new plate.. smaller transfer wedges may be a little more difficult to work with at first, but they are less likely to carry contamination to the new plate. So try practicing taking smaller transfers each time until you're able to easily transfer a piece that is about the size of a grain of rice.

Wait for the mycelium to grow out.. it will be very thin and whispy, and it will grow out toward the edges much quicker than any contamination that tags along.

When the mycelium has grown out enough that I feel confident it has outgrown the contamination (its a good idea to let it at least half way to the edge, or all the way to the edge for more piece of mind), I take a few transfers from the water agar plate and move them to nutritional plates.

Next step is easy: watch your new plate as the mycelium grows out clean!

Edit:
Here's a video I found that discusses a couple applications for water agar that I didn't mention



Pictures added 8/28:

Here are some pictures to show you guys the growth on a few plates.. they aren't the best pictures,  but you should be able to see the leading edge to get an idea of the speed of growth.

Plate stared 8/10 (18 days ago):



8/11 (17 days ago):



8/12 (16 days ago):




Agar to grain comparison
Water Agar vs Nutrient Agar

10/28:
I'm on a mission to start using up some of my ridiculously large plate collection.. So, I decided to do a side-by-side, plate to grain, comparison using 2 fully grown out Phobos plates.. One plate is water agar, the other is MYA. I decided to do 3 jars from each plate.

Quick picture before getting to work:


Here are the plates I'll be using


Plate 1 (MYA- cut into 3 wedges, leaving the center and outer ring behind:


Used plate 1 to inoculate 3 jars of oats:


Here is plate 2 (WA- cut in a similar pattern to plate 1)


Used plate 2 to inoculate the other 3 oat jars:


I plan to post updates comparing any differences in the length of time it takes for the mycelium to jump off, how long it takes the jars to colonize, etc...

11/3

The WA plates are finally starting to jump off to the grain a little bit.. they are definitely running a few days behind the jars that were inoculated using nutrient agar.. like I said in a previous post, this could be due to the age of the plates.. it is a possibility that the mycelium was in a dormant state, due to the fact that they're around 3 months old.. I still plan to run another test with fresh plates to see if the results differ from this batch.

Enigma (WA)


Phobos (WA)


Phobos (MYA)



Edited by Rotnpins (11/03/22 10:56 AM)


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Re: Let's talk about Water Agar!! 💧 [Re: san pedro guy]
    #27900540 - 08/12/22 11:52 AM (1 year, 5 months ago)

Quote:

san pedro guy said:
nice!

have you tried transferring some bacteria and myc on the same plate?

Im also curious if the water agar would work better or worse for sandwhich agar





Yes.. I've transferred mycelium that's grown through bacteria and its enmeshed. It works really well for bacterial mycelium..

I actually just started a bunch of new water agar transfers yesterday so I can update with some pictures showing the process and how the nutritional plates look before and after cleaning up with water agar.. so hopefully I'll be updating in a few days with pictures showing the process


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Re: Let's talk about Water Agar!! 💧 [Re: wild_aussie] * 1
    #27900598 - 08/12/22 12:44 PM (1 year, 5 months ago)

Quote:

wild_aussie said:
Great write up just really getting into agar myself and have had issues with slow growth where bacteria had a chance to take hold can't wait to try this.
:myman:




For sure! I've only done a couple test runs so far, but the results have been really good... I have a feeling that this will be a game changer.. I'm keeping water agar plates on standby so I've got a reliable tool ready whenever I've got a contaminated plate that needs some extra TLC.

If anyone gives it a try, I'd really appreciate it if you shared your experiences and photos in this thread so everyone can see the results from multiple people :mushroom2:


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Re: Let's talk about Water Agar!! 💧 [Re: san pedro guy]
    #27900645 - 08/12/22 01:17 PM (1 year, 5 months ago)

Quote:

san pedro guy said:
I meant more like transfer myc on one part of the plate then transfer some bacteria to the other side (or a separate plate) to see how each responds to the nutritional deprivation

either way i’ll be following; good shit rotn :thumbup:





That's a good idea 💡 I don't have any plates with bacterial colonies that haven't been overtaken by mycelium at the moment, but I'm sure I'll come across some soon and give it a try and post the results


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Re: Let's talk about Water Agar!! 💧 [Re: zilker]
    #27901235 - 08/12/22 07:23 PM (1 year, 5 months ago)

Quote:

zilker said:
Quote:

That's a good idea 💡 I don't have any plates with bacterial colonies that haven't been overtaken by mycelium at the moment, but I'm sure I'll come across some soon and give it a try and post the results




Shouldn't be too hard to find bacteria.  Grab your kitchen sponge, open one of your plates, touch the corner of the sponge to the agar, and seal her back up.  Or, I can send you a complimentary plate in the name of science!  Lol.

On a more serious note, I see huge potential for this as being a 2 transfer until clean, opposed to a 3 transfer until clean.  That will literally save days, weeks, and for some of you old timers, years in transfer/wait time.

I'll definitely be following this post in the future.  Nice job, Rotnpins!





:laugh2: true.. but like you said, it's not too difficult to find bacteria. I'm sure I'll have some on at least one of my germ plates soon :lol:


Exactly 💯  I completely agree...my goal, over time, is to consistently reduce the number of transfers needed, even when starting with heavy contamination.


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Re: Let's talk about Water Agar!! 💧 [Re: Grind365]
    #27901618 - 08/13/22 05:48 AM (1 year, 5 months ago)

Quote:

Baba Yaga said:
Cool stuff, will try this with clone tissue, will be interesting to see if that will
make the washing wish H2O2 or bleach obsolete.

:thumbup::thumbup::thumbup::thumbup::thumbup:




Awesome! You should post your results here so we can see how it goes


Quote:

Grind365 said:
So would you use this all the time or only when you need to clean up a culture?




I would just do this for clean up..

If you're wanting a bit quicker growth on your nutritional plates, you can always try out a low nutrient level.


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Re: Let's talk about Water Agar!! 💧 [Re: Grind365]
    #27902339 - 08/13/22 07:19 PM (1 year, 5 months ago)

Quote:

Awestruck said:
That's a really cool, novel idea. Sounds good. Thanks for sharing the tek!





Quote:

Grind365 said:
Thanks Rotn.





No problem, glad you guys are finding the information useful/interesting :mushroom2:


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Re: Let's talk about Water Agar!! 💧 [Re: baz]
    #27902778 - 08/14/22 07:19 AM (1 year, 5 months ago)

Quote:

baz said:
Can you germinate spores on water agar?




I haven't tried to do it, but I may toss some spores on a plate later and post the results 🤔 

Personally, I like to use plates that have slightly less nutes and agar than my normal plates when making plates specifically for germination... using less agar will make the agar softer and there will be more water to help hydrate the spores.. using less nutrients will help the plate colonize faster, giving you quick growth to transfer to a new plate.

Sometimes you'll see spores germinate in a non nutritional syringe, so I think its possible the plate would germinate.. however I suspect that anything that does germinate will be thin and whispy and hard to distinguish what, if anything, would be worth transferring.

My recommendation for now would be to use nutrient plates for germination.. this will give the spores energy to germinate, and the mycelium will probably be easier to identify.. but I'll post my findings after I give it a try.


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Re: Let's talk about Water Agar!! 💧 [Re: nosf3r4tu]
    #27902965 - 08/14/22 09:50 AM (1 year, 5 months ago)

Quote:

nosf3r4tu said:
I'm a lazy fuck and I just flip the bacterial plates and take a small sample from the other side. Myc grows through the agar, bacteria doesn't.




Interesting.. I'll give that a try sometime and see how it turns out.. hadn't thought of that :super:


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Re: Let's talk about Water Agar!! 💧 [Re: zilker]
    #27910001 - 08/19/22 05:23 PM (1 year, 5 months ago)

Awesome job man! Love the slide pictures to go along with it.. thanks for posting your journey :super:

I've had success taking a clean transfer back to MYA after just a few days if it's decently clean to start with... but if you can see contams tagging along, wait until the WA plate is almost grown out and then take a transfer from the leading edge just to be safe...

or grab a transfer or 2 right away, and hold on to the plate just incase they aren't clean.. by the time you see if the transfer was clean, the WA plate will be grown out a lot further from the contams


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Re: Let's talk about Water Agar!! 💧 [Re: Rotnpins]
    #27910025 - 08/19/22 05:43 PM (1 year, 5 months ago)

Quote:

p9hu7 said:
Water agar is a great tool to have in the toolbox:thumbup:




For sure! I'm surprised how little it's been discussed around the forums..

I had heard it mentioned a couple times, but never with any results posted or certainty that it works..

I wish I had taken more pictures of some of my worst plates so I could share more of the results...

I just transfered some APE today that started out really bacterial and I thought it was a "trash plate". I didn't know about water agar when I was first working in this plate, so the mycelium grew through the bacteria and sat on this plate for a month before transferring to water agar.





This was the plate after transferring from water agar (I only left it on WA a few days, I didn't let it grow out very far, and it looks like it still did its job)



Note: I'm still learning, and haven't tested it on grain yet.. it still looks a bit disorganized, which is why I still did another transfer before doing a test jar.. I'll post the results once I have them


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Re: Let's talk about Water Agar!! 💧 [Re: Psicomb]
    #27913601 - 08/22/22 12:17 PM (1 year, 5 months ago)

That's a brilliant idea 💡  I was planning on making master slants of a couple cultures real soon.. I was going to do 2 slants of each (I always backup my backups :lol: ) but I think I'll probably end up doing 1 set of slants and one set of micro centrifuge tubes with water agar suspended in sterilized distilled water)..

I really like the idea of using the water agar for water suspension vs a wedge from a nutrient plate :super:


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Re: Let's talk about Water Agar!! 💧 [Re: rumfor69] * 1
    #27913628 - 08/22/22 12:40 PM (1 year, 5 months ago)

Quote:

rumfor69 said:
I'm curious if it would starve without any nutrients in long term storage.





I'll probably make a few extras and try reviving them in intervals (3mos 6mos, 1year, etc) and do a write up sharing the results as I go..


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Re: Let's talk about Water Agar!! 💧 [Re: Rotnpins] * 2
    #27913642 - 08/22/22 12:48 PM (1 year, 5 months ago)

Cool.. I'll do it then.. I already planned on trying it out, so it's really no extra effort to make a few extras and test the viability.

Just to make it a little more interesting,  I'll do a set in cold storage and a set at room temp to see if it makes a difference


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Re: Let's talk about Water Agar!! 💧 [Re: Psicomb] * 1
    #27914083 - 08/22/22 06:37 PM (1 year, 5 months ago)

In the name of

:havesomescience:


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Re: Let's talk about Water Agar!! 💧 [Re: Rotnpins]
    #27914123 - 08/22/22 07:03 PM (1 year, 5 months ago)

I haven't done any slants yet.. but I'm planning on retiring my TW clone culture for a while after I see how the current batch turns out, or at least making a master backup.. so this is really good timing to bring this up. It won't take much work to do a quick transfer to water agar while I'm waiting for it to fruit and then try it both ways (slant and water suspension)

Very cool that this write up is starting to generate some creativity.. :rockon:  the only reason we grow the way we do now is because of the community sharing and testing new ideas together :mushroom2:


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Re: Let's talk about Water Agar!! 💧 [Re: Rotnpins]
    #27917108 - 08/24/22 06:18 PM (1 year, 5 months ago)

I found an interesting video for you guys..

He talks about a couple of the applications for water agar that I hadn't mentioned..



Check it out and let me know what you think..


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Re: Let's talk about Water Agar!! 💧 [Re: HILLBILLY OUTLAW]
    #27917162 - 08/24/22 06:56 PM (1 year, 5 months ago)

Quote:

HILLBILLY OUTLAW said:
Great vid addition. That guy has some pretty good vids. Watched a lot of them. I’m defiantly prepping some on my next pour session. You got me. Hook line and stinker!!!





Yeah.. I didn't watch ANY YouTube videos on mycology up until very recently.. I heard that there was a lot of misinformation, so I only learned from information on the shroomery until I thought I had enough of a foundation to weed out the bullshit..

I haven't found a lot of videos I like, but Yoshi Amano and Gary from Fresh From the Farm Fungi have really stood out to me for having solid videos..

I just stumbled on this one today when watching some of Gary's videos and found it pretty interesting.

I really liked how he used it to get a sample straight from some substrate.. I don't think I would have thought of doing that.


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Re: Let's talk about Water Agar!! 💧 [Re: Grind365]
    #27917354 - 08/24/22 09:48 PM (1 year, 5 months ago)

We had a discussion the other day (I believe it was in the cultivation general discussion thread) about all the different myco words that are commonly pronounced different ways:

Albino, Psilocybe, Agar, Cubensis, etc..

I personally pronounce it the same way you do, but there seems to be a lot of variation when it comes to some common terms used in mycology :shrug:


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Re: Let's talk about Water Agar!! 💧 [Re: Grind365]
    #27917362 - 08/24/22 09:51 PM (1 year, 5 months ago)

Quote:

Grind365 said:

Edit: I also noticed he said more nutritious agar gives fast vigorous growth I know the opposite to be true what gives?




In comparison to water agar, it's definitely a lot quicker and more vigorous. However this works to out advantage because it still moves faster than contamination.


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