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Catattack Some Agar (again)


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#341 CatsAndBats

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Posted 05 November 2016 - 01:00 PM

Found this today:

 

https://mycotopia.ne...a-weight-scale/

 

might be of use to somebody



#342 CatsAndBats

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Posted 09 November 2016 - 10:35 AM

Okay, so everything exploded! I mean all three cultures that I germinated or transferred on Nov. 1st are showing good growth. So that's the "albino" B+ from spore, the "albino" redboys from spore, and although not as surprisingly the malabar rhizo that I've come to love. The spore germinated myc will be under my watchful eye looking for any hitch-hikers.

 

Today's recipe is:

12g agar

5g sucrose

red stain

(no catattives today)

 

The purpose of today's recipe is to see if I can force rhizomorphic growth with "low" nutrition, which is also why I left the catattives out.

_______________________________________________________________________________

 

I get quite a few PM's about various agar questions and I wanted to let anybody considering getting into agar know, that it is not that difficult and once one dives into it, one will be rewarded ten fold. All of these recipes that I post or reference are not necessary , I do them for fun or for specific nerdy purposes.

 

My first successful agar recipe was dry dog food and agar. If you are going to make your own, all you need is agar and sugar and water. If making your own agar is not for you, by all means, buy pre-poured plates from a reputable source!

 

Agar allows us to see what is actually going on and is a GREAT way to germinate spores. It allows a grower to take mycelium and train it, clean it, manipulate it, test it, etc.. It's easy!

 

Basically, dive right in, the water is purrfect!


Edited by catattack, 09 November 2016 - 10:35 AM.

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#343 mycolangelo.4HO

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Posted 09 November 2016 - 12:27 PM

Reminds me of the American Dad episode where Roger opens a bar. Dive on in.
Don't know how to add a clip and I'm working but I thought someone might be able to do it. But I agree I never really had much luck with lc's so I went to agar and haven't looked back. I buy 500g pda and it's very cheap when you think about how much agar that will make, I don't use plates I just use jars. But I love how you can take a dirty culture and clean it up very quickly. I was a bit hesitant cuz it seemed overly complicated, but it's really easy.

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#344 CatsAndBats

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Posted 09 November 2016 - 12:38 PM

Reminds me of the American Dad episode where Roger opens a bar. Dive on in.
 

 

Are we related? Roger Smith is my spirit animal.


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#345 mycolangelo.4HO

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Posted 09 November 2016 - 02:22 PM

Ah you're spirit animal that's some good shit (dive on in),

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#346 CatsAndBats

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Posted 09 November 2016 - 03:47 PM

@peacefrog, just checked the pans, they took to the grain!


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#347 CatsAndBats

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Posted 10 November 2016 - 09:54 PM

So here is the above par malabar culture that I pulled off of that real aggressive rhizo mentioned in a recent post

 

( https://mycotopia.ne...in/?p=1293137).

 

I'm going to start five more transfers.

 

This is the homemade agar that I made last that is recorded on this thread. The jars are upside down because there is a little extra moisture that I want to wipe out. This will be achieved in an aseptic fashion due to the fact that I can quickly and mindfully remove the lid and place the jar rim side down onto clorox disinfectant  wipes that I line the bottom of the SAB with. Since they are facing down no airborne particles can fall in.

 

post-147940-0-65188800-1478832241.jpg

 

post-147940-0-12200800-1478832147.jpg

 

 

 

post-147940-0-71624900-1478832174.jpg

I keep my two favorite tools in a jar of ethanol. A dental scraper and  tissue sampling forceps that have little teeth to grab flesh from a fruit body or agar plug.

 

I got everything into the SAB thoroughly wiped down and sprayed all sides with lysol kitchen spray, only wiping the surface that I peer through, letting the rest remain wet to catch any airbornes that are left. I then walk away from it for ~15min to let the dust settle.

 

Go in with gloved hands and take my sectors!

 

 

agar1.jpg

agar SAB.jpg

Attached Thumbnails

  • agar mal.jpg

Edited by catattack, 10 November 2016 - 10:02 PM.

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#348 CatsAndBats

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Posted 11 November 2016 - 08:29 AM

Correction

 

This is the recipe that I used in those grey-ish jars in the previous post:

https://mycotopia.ne...gain/?p=1293192

 

This is the red recipe that's low nutrients to force rhizo that will be the next transfer:

https://mycotopia.ne...gain/?p=1294122

 

 

Oh and here are two of my favorite aforementioned agar tools. Left is the tissue sampling tweezers/forceps and right is the dental scraper. The scraper can be used as a cutting tool if used backwards and is dull enough to stab a piece of agar and move it without it sliding right off like a scalpel.

 

 post-147940-0-16028900-1478874672.jpg

Attached Thumbnails

  • agar_tools.jpg

Edited by catattack, 11 November 2016 - 09:32 AM.

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#349 CatsAndBats

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Posted 13 November 2016 - 11:44 AM

@peacefrog, here's where the new pan grow is:

https://mycotopia.ne...ome-pans-again/



#350 CatsAndBats

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Posted 13 November 2016 - 05:13 PM

 post-147940-0-99299800-1479074872.jpg

 

Here's my ladling technique for getting the nutrients evenly distributed. Now I simmer in the jar to help distribute the nutes already, but I also ladle from the bottom for each run and this ladle gives me the purrfect amount for my little jars.

Attached Thumbnails

  • agar ladel.jpg

Edited by catattack, 13 November 2016 - 05:14 PM.


#351 HrVanker

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Posted 13 November 2016 - 06:34 PM

Reminds me of the American Dad episode where Roger opens a bar. Dive on in.


[Direct Link]





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#352 mycolangelo.4HO

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Posted 14 November 2016 - 12:54 PM

Thank you, great way to start the day (dive on in)

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#353 CatsAndBats

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Posted 15 November 2016 - 08:34 AM

post-147940-0-86885900-1479216403.jpg 

 

So I had several jars started from spore (b+ leucistic and redboy leucistic) that showed a ton of tomentose growth. I want to force rhizomorphic growth so I transferred to the half nutritious red agar pictured in a previous post. In about 5-7 days I should hopefully see more rhizomorphic growth and if not I'll lower the nutrients again.

 

I'll probably make low nute agar and regular nute agar from here on out and have them on hand depending on what I want the myc to do.

Attached Thumbnails

  • tomentose.jpg

Edited by catattack, 15 November 2016 - 08:34 AM.


#354 JustAnEyedea

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Posted 17 November 2016 - 02:29 PM

I have also dove back into agar after recently finding a couple plates of my ksss! I only did a few no pour agar jars to make sure my recipe was correct. A few transfers actually sank right into the agar lol. Those won't work! I'm assuming I didn't mix it up enough to begin with. I used a PDA premix that I have probably possessed for way to long without having done anything with it.

I used 25gms PDA
9gms dextrose
1grm bacto peptone ( not sure if this it's necessary)
I'm hoping to stretch this out to multiple cultures and share them with my fellow topiates.tmp_5630-1117161215-600x80025225120.jpg

This one sank into the agartmp_5630-1117161214a-600x800-409220205.jpg

These ones seem finetmp_5630-1117161214b-600x8001693939356.jpg

tmp_5630-1117161214-600x800-1001431716.jpg

It's going to take me a while to read all of this thread, so I thought I'd post first and add to this monster!
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#355 CatsAndBats

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Posted 17 November 2016 - 09:07 PM

 post-147940-0-48767100-1479434773.jpg

 

Malabars continuing to look better. Nice radial growth.

 

I want to see it run the low nutrient agar I made up.

Attached Thumbnails

  • malabars.jpg

Edited by catattack, 17 November 2016 - 09:09 PM.

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#356 Microbe

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Posted 17 November 2016 - 10:41 PM

post-147940-0-48767100-1479434773.jpg

Malabars continuing to look better. Nice radial growth.

I want to see it run the low nutrient agar I made up.

That is uniform growth and pretty dang close to a isolate. Dont let the lack of that roping rhizo fool you. That is considered rhizomorphic growth btw which you know that.

ecc4fed1d8a7d5608dcfecc4f422a0f4.jpg


I want to talk about this. Where the arrow is pointing is the filaments of hyphae branching out ahead of the mycelia colony. This is where the chemical signals are happening but what i really want to say is that is where you want to transfer from when trying to isolate a monokaryon but a little sooner and as soon as you see that haze. Obviously thats not a monokaryon but you will see the same thing from a monospore germination but you have to grab it as soon as you see the first sign and from the edge.

Now something else i have seen and never even thought of this, i have seen monokaryotic mycelium left behind. Where mating has taken place and it is now dikaryon growing out with the monokaryon left in the back field so to speak, you could grab it from there if that ever happened and with confidence. You would need to do it before the plate became colonized because the dikaryon will most certainly will start growing back over the monokaryon.

You know what im saying homeboy?

Edited by Microbe77, 17 November 2016 - 10:42 PM.

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#357 Microbe

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Posted 17 November 2016 - 10:43 PM

post-147940-0-86885900-1479216403.jpg

So I had several jars started from spore (b+ leucistic and redboy leucistic) that showed a ton of tomentose growth. I want to force rhizomorphic growth so I transferred to the half nutritious red agar pictured in a previous post. In about 5-7 days I should hopefully see more rhizomorphic growth and if not I'll lower the nutrients again.

I'll probably make low nute agar and regular nute agar from here on out and have them on hand depending on what I want the myc to do.

Dude im sending you some plates. Im sick of that silliness! How many you want?
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#358 CatsAndBats

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Posted 17 November 2016 - 11:11 PM

 

post-147940-0-86885900-1479216403.jpg

So I had several jars started from spore (b+ leucistic and redboy leucistic) that showed a ton of tomentose growth. I want to force rhizomorphic growth so I transferred to the half nutritious red agar pictured in a previous post. In about 5-7 days I should hopefully see more rhizomorphic growth and if not I'll lower the nutrients again.

I'll probably make low nute agar and regular nute agar from here on out and have them on hand depending on what I want the myc to do.

Dude im sending you some plates. Im sick of that silliness! How many you want?

 

 

I love my jars. I can see everything I need to.

 

Good looking out on the other shot. I love the growth.



#359 Microbe

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Posted 17 November 2016 - 11:16 PM

post-147940-0-86885900-1479216403.jpg

So I had several jars started from spore (b+ leucistic and redboy leucistic) that showed a ton of tomentose growth. I want to force rhizomorphic growth so I transferred to the half nutritious red agar pictured in a previous post. In about 5-7 days I should hopefully see more rhizomorphic growth and if not I'll lower the nutrients again.

I'll probably make low nute agar and regular nute agar from here on out and have them on hand depending on what I want the myc to do.

Dude im sending you some plates. Im sick of that silliness! How many you want?
I love my jars. I can see everything I need to.

Good looking out on the other shot. I love the growth.
So a 3 dozen? I will bring them with me if i ever go visit my family round yo neck of da woods....

Edited by Microbe77, 17 November 2016 - 11:17 PM.

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#360 Yumbum

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Posted 20 November 2016 - 04:10 PM

Hi guys, so Im gonna jump in here and hopefully some more experienced eyes can tell me what Im looking at and for here in these Pan Cyanenscens Hawaiians.

All of these dishes are second generation. Here's the thing I don't get. I see the white fluffy stuff and then the mycelium sort of goes transparent. Is this the best growth to take to isolate further, the almost see thru stuff at the edges? Or is this all like that coz it's drying out? I've definitely noticed that some of the agar is drying out around the edges, so I'm gonna pour thicker next time, but in the meantime, does this make sense to anyone? Also notice how some of the fluffier stuff is kinda raggedy looking? Any thoughts on that as well? 

2.jpg 1.jpg agar.jpg


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