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Agar blues


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#1 Salty117

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Posted 02 July 2021 - 07:29 AM

I mentioned in a thread somewhere about mycelium looking strange, sort of gelatinous on my agar. I have several cups that are looking like this but I'm just not sure where I went wrong. I was told the rule of thumb is 2g agar agar per 100ml of water but I thought I would post up a picture of what exactly I'm looking at in hopes someone might be able to help me improve. Thank you for your time

 

210410231_599795000985182_5348251383450326396_n.jpg


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#2 Sidestreet

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Posted 03 July 2021 - 11:37 AM

2 g per 100 mL sounds right to me.  What's the rest of your recipe?


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#3 Salty117

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Posted 03 July 2021 - 02:37 PM

10g ELME

8g Peptone

16g Dextrose to 500ml water. with the 10g of agar agar


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#4 Salty117

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Posted 03 July 2021 - 08:26 PM

I mix it in a 1000ml flask with a magnetic stir bar on a stir plate, add my dry ingredients first and I boil water in a tea kettle to a rolling boil before adding it to the flask. I've been using tap water (city) but I'm probably going to start using the filtered water from our brita jug since the house is old and so is the plumbing.

The only other agar I've made was that same recipe but also including 1g of powdered activated charcoal per 500ml batch but that seems to have a little less of a gel-like effect with the one transfer I made to one with a copelandia specimen that thrived from LC to one of the beige agar plates.

 

212168812_559979241803147_7659410383255331192_n.jpg

 

211511876_307223091095400_6402289941534447696_n.jpg

 

I feel like I have read and watched a lot of methods of how to work with agar in a SAB or in front of a flow hood and when the time comes that I'm starting to make my cuts and do the work I forget some of the little things like how thin to pour my cups or how big to cut my transfers. I'm sure I'll get the hang of those things in time though  :smile:


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#5 Sidestreet

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Posted 04 July 2021 - 07:34 AM

10 g LME and 16 g dextrose?  Is that too much food/sugar? 


Edited by Sidestreet, 04 July 2021 - 07:34 AM.

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#6 Salty117

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Posted 04 July 2021 - 10:12 AM

Perhaps? I was referencing JOC's preferred agar recipe mentioned in the Official Copelandia thread but at the time I made this batch I had forgotten to get potato flakes and decided to go with ELME.. I can see how the two wouldn't have the same "sugar" content but I didn't have an idea of how much sugar contant the ELME would provide


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#7 Salty117

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Posted 04 July 2021 - 10:19 AM

Should I cut the dextrose and see how it goes for my next batch? I used the same amount of ELME and Dextrose in the Black Agar and the one that seemed to make it through without getting contams of some sort seems to look good which is why I wasn't sure if it was the dextrose that was making the other cups look funny or if I had too much water or not enough agar agar.

 

If the water/agar agar I used is normal then I'll try whipping up a batch without the Dextrose and see how that goes


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#8 Salty117

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Posted 04 July 2021 - 11:20 AM

I have been told it's safe to presume Malt Extract to be upto 50-70% sugar so I suppose with 16g dextrose to 500ml of boiling water would be way too much. Going to simplify things and work my way up from there


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#9 Oldpunk

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Posted 04 July 2021 - 11:09 PM

I've only made a few batches of agar myself, but I didn't use any sugar with malt extract.

Mine was 10g malt 10g agar 0.8g crushed nutritional yeast. To 500 ml

I tried potato flakes first and felt I had too much sugar cuz I got real thick fuzzy growth most of the time. But here I am a few months later and all of those fuzzy agar cups grew out just fine (well most of them). I think you've got way too much nutrient in your mix and it's growing in super fast and thick.
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#10 rockyfungus

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Posted 05 July 2021 - 01:12 AM

10g ELME

8g Peptone

16g Dextrose to 500ml water. with the 10g of agar agar

Way too much food. Less is more.

If we are working with 10g of agar. That's 500ml of H20 for the average stiffness. I prefer 8g agar for the extra adhesion.

 

10g of any sugar is the max I do and lately I've been dropping that super low so that myc has to reach for food and get that beautiful rhizo people like. 

Never played with peptone it is def. not needed, unless you are following textbooks. 

If you are making grain just look at the water. So 500g of any grain juice I'll dump to 100g-50g of juice and add back 400g of h2o. Just that nice light amber beer color...

 

The Usual

500ml H20

8g agar

8g malted stuff1

pinch of nutritional yeast or additives (straw, manure, wood, compost)

 

 

1: all your malted starchy stuff (complex sugars?), potato + dextrose (people don't care for this, never tried it), GRAIN WATER


Edited by rockyfungus, 05 July 2021 - 01:21 AM.

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#11 CatsAndBats

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Posted 05 July 2021 - 02:29 AM

I mix it in a 1000ml flask with a magnetic stir bar on a stir plate, add my dry ingredients first and I boil water in a tea kettle to a rolling boil before adding it to the flask. I've been using tap water (city) but I'm probably going to start using the filtered water from our brita jug since the house is old and so is the plumbing.

The only other agar I've made was that same recipe but also including 1g of powdered activated charcoal per 500ml batch but that seems to have a little less of a gel-like effect with the one transfer I made to one with a copelandia specimen that thrived from LC to one of the beige agar plates.

 

 

 

attachicon.gif211511876_307223091095400_6402289941534447696_n.jpg

 

I feel like I have read and watched a lot of methods of how to work with agar in a SAB or in front of a flow hood and when the time comes that I'm starting to make my cuts and do the work I forget some of the little things like how thin to pour my cups or how big to cut my transfers. I'm sure I'll get the hang of those things in time though  :smile:

 

Tap water is fine, unless you have really really shitty tap water. Don't overthink it (I know that it's hard to avoid thinking too much, but just try not to).

 

10 g LME and 16 g dextrose?  Is that too much food/sugar? 

 

IMO no, I mean it's a waste of ingredients, but the myc will still colonize it IME.

 

 

10g ELME

8g Peptone

16g Dextrose to 500ml water. with the 10g of agar agar

Way too much food. Less is more.

If we are working with 10g of agar. That's 500ml of H20 for the average stiffness. I prefer 8g agar for the extra adhesion.

 

10g of any sugar is the max I do and lately I've been dropping that super low so that myc has to reach for food and get that beautiful rhizo people like. 

Never played with peptone it is def. not needed, unless you are following textbooks. 

If you are making grain just look at the water. So 500g of any grain juice I'll dump to 100g-50g of juice and add back 400g of h2o. Just that nice light amber beer color...

 

The Usual

500ml H20

8g agar

8g malted stuff1

pinch of nutritional yeast or additives (straw, manure, wood, compost)

 

 

1: all your malted starchy stuff (complex sugars?), potato + dextrose (people don't care for this, never tried it), GRAIN WATER

 

I have to respectfully disagree. In my experience, the nutrition amount in and of itself is shouldn't be detrimental to the mycelium. That said, it's way too much nutrition :tongue:

 

Salty, I think that your problem is the dextrose. Dextrose can set weird, even if one uses powdered dextrose, it'll definitely set weird if one uses dextrose syrup.

 

Do yourself a favor, do a batch of agar with the following ratios:

 

agar: 10g

brf 10g

500ml h2o

 

and go from there, I bet you'll have success

 

If you'd like to read more about my theories/propaganda, go here:

 

https://mycotopia.ne...izes-agar-agar/

 

and here:

 

https://mycotopia.ne...k-some-agar-30/


Edited by CatsAndBats, 05 July 2021 - 02:30 AM.

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#12 rockyfungus

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Posted 05 July 2021 - 02:35 AM

No-one said anything about adverse affects of too much food. It's a waste of money plain and simple, I'm frugal. 



#13 Salty117

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Posted 05 July 2021 - 08:18 AM

 

10g ELME

8g Peptone

16g Dextrose to 500ml water. with the 10g of agar agar

Way too much food. Less is more.

If we are working with 10g of agar. That's 500ml of H20 for the average stiffness. I prefer 8g agar for the extra adhesion.

 

10g of any sugar is the max I do and lately I've been dropping that super low so that myc has to reach for food and get that beautiful rhizo people like. 

Never played with peptone it is def. not needed, unless you are following textbooks. 

If you are making grain just look at the water. So 500g of any grain juice I'll dump to 100g-50g of juice and add back 400g of h2o. Just that nice light amber beer color...

 

The Usual

500ml H20

8g agar

8g malted stuff1

pinch of nutritional yeast or additives (straw, manure, wood, compost)

 

 

1: all your malted starchy stuff (complex sugars?), potato + dextrose (people don't care for this, never tried it), GRAIN WATER

 

 

Is the grain water something I would need to use in a batch of agar right when it's available to use or can I store it in a sealed container in the fridge for a couple days if need be without bacteria thriving in it?

 

 

 

 

Salty, I think that your problem is the dextrose. Dextrose can set weird, even if one uses powdered dextrose, it'll definitely set weird if one uses dextrose syrup.

 

Do yourself a favor, do a batch of agar with the following ratios:

 

agar: 10g

brf 10g

500ml h2o

 

and go from there, I bet you'll have success

 

If you'd like to read more about my theories/propaganda, go here:

 

https://mycotopia.ne...izes-agar-agar/

 

and here:

 

https://mycotopia.ne...k-some-agar-30/

 

 

I did use powdered Dextrose in my failed attempt at Agar. I have lots of cups and lots of spores to work with so I'll give this recipe a shot. Before I read this I did make another batch of Agar using 10g ELME and 8g Peptone 10g agar agar to 500ml boiling water so I'm not going to let that go to waste but I plan to make more soon.

 

 

No-one said anything about adverse affects of too much food. It's a waste of money plain and simple, I'm frugal. 

 

In this case the adverse effects I'm seeing is strange gelatinous 'rhizomorphic' growth that is stalling.


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#14 rockyfungus

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Posted 05 July 2021 - 09:27 AM

Used to freeze grain water (broke too much glass), stored in fridge for a week was never an issue, it's getting sterilized so as long as it doesn't ferment probably fine. 


Edited by rockyfungus, 05 July 2021 - 09:27 AM.


#15 Oldpunk

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Posted 05 July 2021 - 09:53 AM

I tried the grain water this time around. Ive only tried 2 cups so far but it seems happy. Some PE and mystery mush sprouted myc nicely.

I don't think I used anything but 10g agar to 500ml grain water left after boiling bird seed.
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#16 Arathu

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Posted 05 July 2021 - 02:36 PM

 

Do yourself a favor, do a batch of agar with the following ratios:

 

agar: 10g

brf 10g

500ml h2o

 

and go from there, I bet you'll have success

 

After all these years....this recipe still works like a charm IME.......

 

I agree....a great starting, and in fact continuing, point....

 

Combined with an antibiotic or two if necessary most germination and clean up work can be accomplished using the above......

 

A


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#17 rockyfungus

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Posted 05 July 2021 - 03:19 PM

 

 

No-one said anything about adverse affects of too much food. It's a waste of money plain and simple, I'm frugal. 

 

In this case the adverse effects I'm seeing is strange gelatinous 'rhizomorphic' growth that is stalling.

 

Sorry was tripping :)

Did ya have control plates. I'm about to go check mine. I've been getting slimed, molded, triched and things I've never seen from some reddit stickers, and the SEASON!

 

But yeah the 10:10:500 is just easy to remember. Go low, lower on agar if you feel you can't keep it on the blade. I invert all my dishes so I can literally stick a sticker small piece on stiffer agar. 10g on T0. 8g T1, and then go up to like 9g on t2. That's all I usually do print and recycle. Maybe clone a nice cluster, or keep a sexy rhizo sector for further isolation. 

Lower on the sugars/food, saving money is fun, experiment with random stuff (pure compost juice is my next project). 


Edited by rockyfungus, 05 July 2021 - 03:21 PM.

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#18 Salty117

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Posted 06 July 2021 - 01:22 PM

 

 

Do yourself a favor, do a batch of agar with the following ratios:

 

agar: 10g

brf 10g

500ml h2o

 

and go from there, I bet you'll have success

 

After all these years....this recipe still works like a charm IME.......

 

I agree....a great starting, and in fact continuing, point....

 

Combined with an antibiotic or two if necessary most germination and clean up work can be accomplished using the above......

 

A

 

 

What antibiotics would you happen to recommend, Arathu? so far I've been recommended Furan-2 fish antibiotics but I haven't gone ahead and gotten it yet so I'm open to recommendations and seeing what my options are


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#19 Salty117

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Posted 27 July 2021 - 12:11 PM

Back with /some/ results, The Huautla water agar was lost to contamination and so was the Tennessee Cubensis BRF agar.

 

As I'm sure it's easy to assume, the water agar took a little over a day and a half longer than the PDA or BRFA to show visible growth, but I am wondering if the spores that germinate to a less nutritional source are the "fittest" since there'd be less encouragement for weaker spore/genetics to thrive? Then the assumed "stronger" genetics get transferred out to more nutritional agar and grow out before the other possibly weaker spores can get to/thrive upon the new and more nutritional food source. Just a curious thought by an aspiring mycofanatic, perhaps someone has already gone through this hypothesis and tested it out lol, it wouldn't surprise me.

 

Wild Tennessee Cubensis  on water agar, made transfers out to some choice PDA cups.

 

Super Orange - Wild Cubensis from Tennessee Water Agar (1) Resized.jpg

 

Super Orange - Wild Cubensis from Tennessee Water Agar (2) Resized.jpg

 

Picked up some 60mmx15mm Borosilicate Petris and I am already loving them. Can't wait to get more!  :biggrin:

 

Black Agar in Borosilicate Petris.jpg

 

Huautla Cubensis on PDA, if there's any more experienced agar heads viewing this my question to you is how close do you get to the source of contamination when you make transfers out from that cup/plate?

 

Huautla Cubensis on PDA Resized.jpg

 

Huautla Cubensis on BRFA - I will definitely give it a couple more runs but needing to keep it on the stir plate when the agar is cooling to keep everything suspendid is slight turn off from this recipe but like I said, I will give it a few more runs maybe even use it for the last transfer before going to grains.

 

Huautla Cubensis on BRFA (1) Resized.jpg

 

Huautla Cubensis on BRFA (3) Resized.jpg


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#20 Salty117

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Posted 27 July 2021 - 12:23 PM

Forgot to add these guys to the last post, some Copelandia Cambodginiensis Sandose on ELME Agar with Charcoal and Peptone.

 

Copelandia Sandose on Black ELME Agar with Peptone.jpg

 

Copelandia Sandose on Black ELME Agar with Peptone1.jpg

 

Edit: Double posted a photo when I meant to include this one here

 

Copelandia Sandose on Black ELME Agar with Peptone2.jpg


Edited by Salty117, 27 July 2021 - 12:52 PM.

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