Paradox
©
Fisana

Jump to content


Photo
* * * * * 4 votes

Catattack some Agar 3.0

artwork friend

  • Please log in to reply
201 replies to this topic

#21 whitethumb

whitethumb

    poisoned beauties

  • OG VIP
  • 940 posts

Donator

Posted 25 January 2017 - 11:25 PM

great write up brother, i feel privileged to be mentioned in the same sentence with people i look up to.
  • CatsAndBats likes this

#22 Arathu

Arathu

    Dirtmaker

  • OG VIP
  • 3,775 posts

Awards Bar:

Posted 26 January 2017 - 12:33 AM

WOW! I'm way outta likes dude! I'm going to enjoy watching this.............and play too..........I have grifola to work on over here in the woods............. :cool:


  • JustAnEyedea likes this

#23 ethnobotanist420

ethnobotanist420

    Welcome to the Machine

  • OG VIP
  • 3,468 posts

Awards Bar:

Posted 26 January 2017 - 01:47 AM

This looks awesome!

Haven't read all the links yet but I'm posting so I don't forget to lol

Great work, Cat!!
  • CatsAndBats likes this

#24 CatsAndBats

CatsAndBats

    /(^ x ^)\

  • OG VIP
  • 7,188 posts

Awards Bar:

Posted 26 January 2017 - 08:58 AM

post-147940-0-19202100-1485435261.jpg

 

This one is for @archersMark. The arrow is pointing at a square of agar that was stripped of it's color by the mycelium metabolizing the nutrition and food coloring, which I've been told is caused by the change in pH. It was a batch of red agar that I made, which seems to lose it's color more readily than say blue or green stains. I grabbed the recipe and my notes on it as I think that they are applicable for any newcomer to agar (please note that the recipe is half nutrition and shouldn't be copied):

 

 

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!

Spoiler alert, I wasn't super pleased with this recipe (half nutrition)it didn't force the rhizo like I'd hoped, but it got the job done, obviously. The strains pictured are "amazon albino" which @coorsmikey and I have been trying to figure out the taxonomy on and pioppinos (black poplars). Either way I am super pleased and excited to fruit out in the coming weeks.

 

_______________________________________________________________

 

 

post-147940-0-49234500-1485437360.jpg

 

This next shot is displaying a couple of 'baller' techniques that I use. Now I do 'no pour', meaning I mix my agar up and PC in jars as opposed to pouring them in a clean area after sterilization. The baller moves are how I treat them in the SAB. I line the bottom of my SAB with clorox/lysol wipes, this keeps my working surface sterile/aseptic and catches airborne particles. I wipe my PC'd jars down with these same type wipes. I have a wipe in each hand as I crack the lids, the entire jar is covered with the wipes, and if there's uneven pressure the wipes filter the air when I break the seal. I then put the jars in upside down which forces any excess moisture into the lid. After all of my tools and jars are inside the SAB and wiped down (I don't wipe the walls of the SAB as I want the walls wet to catch more particulates)I walk away to let the dust settle (very important when in an SAB or glovebox). Now after that I go in with my gloved and sanitized hands, pull the lids off and put the jars on my saturated surface, still upside down, this ensures that any particles that may still be floating around can't fall in. The only time that my jars (and the agar surface) are exposed to air is the 2-3 seconds (if that) that it takes to put the tissue/spores/whatever in.

 

This brings up an important point. Learn to love your SAB/clean area. PRACTICE makes perfect. Do dry runs. Practice all of your movements and work. One will greatly benefit from building muscle memory in one's clean area (mad props to @tvcasualty for impressing this upon me). The minute that one starts getting frustrated or making mistakes, will just lead to more mistakes, building the muscle memory and practicing clean technique will help avoid this.

 

I like to put some good hip hop on and get in the 'zone'. If one has fun while doing clean technique with confidence, one will have more success in my opinion and experience. So smoke what you smoke, crack a special microbrew or pop pills or whatever you do to have fun and have some fun. It helps, trust me.

 

It should be noted that the discoloration pictured in those jars are some additives that sank to the bottom, the surface of the agar where the magic happens is pristine.

 

So "DIVE ON IN" kids.

Attached Thumbnails

  • agar2.jpg
  • agar.jpg

Edited by catattack, 26 January 2017 - 09:23 AM.

  • Seeker2be, archersmark, Gnomesayin and 1 other like this

#25 CatsAndBats

CatsAndBats

    /(^ x ^)\

  • OG VIP
  • 7,188 posts

Awards Bar:

Posted 26 January 2017 - 09:17 AM


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

Pictured are two of my main tools that I use, not pictured are my disposable scalpels. I keep these submerged in a jar with 90% alcohol. I dip the handle side in first and then place them in correctly to sit in the alcohol until use. I use 90% for this because it kills unwanted microbes "harder" and evaporates faster than 70% which means they dry quicker before they touch the myc. If I were using alcohol anywhere other than inside my SAB I'd use 70% because it evaporates more slowly, keeping things aseptic for longer. GNOMESAYING?!


Edited by catattack, 26 January 2017 - 09:55 AM.

  • archersmark and Gnomesayin like this

#26 CatsAndBats

CatsAndBats

    /(^ x ^)\

  • OG VIP
  • 7,188 posts

Awards Bar:

Posted 26 January 2017 - 09:52 AM

I have said this before but i dont think many know how much time is involved to lay down a structured thread like this. Your not getting paid to do this, your doing this out of your intense passion to teach and help evolve the cultivator of any skill set or tenor. It is very educational and easy to follow along. Perhaps now that i have nothing but time on my hands i may assist in these teachings. Again brother excellent work and thank your putting together a valuable resource for those who want to learn about the backbone of the hobby. However im deducting a few points and grading this a A-. You know why dont you? I bet you do......

Sent from my LGLS992 using Tapatalk

 

Outta 'likes', thank you my brother. You were instrumental on my last agar thread!


  • Microbe likes this

#27 Arathu

Arathu

    Dirtmaker

  • OG VIP
  • 3,775 posts

Awards Bar:

Posted 26 January 2017 - 10:54 AM

That's exactly how I do it too. My agar gets made up and poured into the jelly jars then I sterilized in the jars. Thanks for the info on metabolizing food dye..........I did not know that......

 

Pay attention class there is much to learn! 

 

Great job Cat!

 

A


  • archersmark, Microbe and CatsAndBats like this

#28 archersmark

archersmark

    Mycophage

  • Free Member
  • 126 posts

Posted 26 January 2017 - 11:26 AM

I might have to find a dental scraper and give it a try.  I find myself cutting with my scalpel and piercing the agar to get it to move.  Where you do find 90% alcohol?  I was wondering what hair stylists use, don't they submerge their tools in a blue alcohol bath until ready to use? 


  • CatsAndBats, JustAnEyedea and Oam like this

#29 CatsAndBats

CatsAndBats

    /(^ x ^)\

  • OG VIP
  • 7,188 posts

Awards Bar:

Posted 26 January 2017 - 11:32 AM

I might have to find a dental scraper and give it a try.  I find myself cutting with my scalpel and piercing the agar to get it to move.  Where you do find 90% alcohol?  I was wondering what hair stylists use, don't they submerge their tools in a blue alcohol bath until ready to use? 

 

Any well stocked pharmacy will have 70% & 90%, target does.

 

The stuff the barbers use is "barbicide" and has an anti-fungal in it so no bueno.

 

https://www.barbicid...de-concentrate/


  • archersmark and JustAnEyedea like this

#30 archersmark

archersmark

    Mycophage

  • Free Member
  • 126 posts

Posted 26 January 2017 - 12:19 PM

 

I might have to find a dental scraper and give it a try.  I find myself cutting with my scalpel and piercing the agar to get it to move.  Where you do find 90% alcohol?  I was wondering what hair stylists use, don't they submerge their tools in a blue alcohol bath until ready to use? 

 

Any well stocked pharmacy will have 70% & 90%, target does.

 

The stuff the barbers use is "barbicide" and has an anti-fungal in it so no bueno.

 

https://www.barbicid...de-concentrate/

 

And this ^^ is exactly why I have started to "think out loud" i.e. mention it to the forum so I learn why that shower thought I had is not such a great idea after all.  Thanks for saving me a whole bunch of agar failures before they happened :)


  • Arathu, JustAnEyedea and burncitymyco like this

#31 CatsAndBats

CatsAndBats

    /(^ x ^)\

  • OG VIP
  • 7,188 posts

Awards Bar:

Posted 26 January 2017 - 01:15 PM

That's literally the point of this thread. Anytime my brother!
  • archersmark and JustAnEyedea like this

#32 Nsnail

Nsnail

    Mycophiliac

  • Free Member
  • 70 posts

Posted 26 January 2017 - 01:27 PM

Have you given any thought to lazlos salt antiseptic teks where he adds salt to agar or soaks tissue / spores in a saline solution before transferring to agar


Sent from my iPhone using Tapatalk

#33 CatsAndBats

CatsAndBats

    /(^ x ^)\

  • OG VIP
  • 7,188 posts

Awards Bar:

Posted 26 January 2017 - 01:37 PM

Have you given any thought to lazlos salt antiseptic teks where he adds salt to agar or soaks tissue / spores in a saline solution before transferring to agar


Sent from my iPhone using Tapatalk

I have not. Got links? That's actually purrfect as the next major installment here will be cloning work!

Edited by catattack, 26 January 2017 - 01:38 PM.

  • Arathu likes this

#34 salviamycelium

salviamycelium

    Mycophage

  • Free Member
  • 192 posts

Posted 26 January 2017 - 01:45 PM

Nice. I'll spy on this thread.


  • CatsAndBats likes this

#35 Nsnail

Nsnail

    Mycophiliac

  • Free Member
  • 70 posts

Posted 26 January 2017 - 01:50 PM

Have you given any thought to lazlos salt antiseptic teks where he adds salt to agar or soaks tissue / spores in a saline solution before transferring to agar


Sent from my iPhone using Tapatalk

I have not. Got links? That's actually purrfect as the next major installment here will be cloning work!
here ya go https://mycotopia.ne...ing-antiseptic/

I tried soaking some portabella tissue but no signs of growth yet
  • CatsAndBats and JustAnEyedea like this

#36 JustAnEyedea

JustAnEyedea

    Destroy Your Television

  • Black VIP
  • 536 posts

Donator


Awards Bar:

Posted 26 January 2017 - 03:46 PM

Looking good everyone! I'm excited for this thread, as I'm continuing to improve my agar skills.

My recipe:
20gms PDA/25lbs MEA per 500 mil or water
(I switch between PDA and MEA to keep the myc interested when doing isolates)
.5gms pulverized oyster shell
1-2gms bacto peptone
Sometimes a pinch of volcanic ash from Mt St Helens
Any food coloring I Like. I prefer black or blue.
Pc for 20-30 minutes at the usual 15psi
Let cool overnight, of course.
Do my transfers, and pray to the Gods

I do not use petries. I use a no pour tek with wide mouth half pints. I've never had good luck with dishes, but as soon as I moved to i jars with the tek I saw a significant decrease in contaminates.

I have a 4x3 hepa flow hood I work in front of in my basement. That helps! I have never tried doing agar work in an SAB but I expect it would be a bitch. I'l

I'm looking forward to seeing where this thread is going, and the contributions by myself and others.

Thanks for another awesome thread, Cat!

Edited by JustAnEyedea, 26 January 2017 - 03:50 PM.

  • CatsAndBats, Gnomesayin and Nsnail like this

#37 Arathu

Arathu

    Dirtmaker

  • OG VIP
  • 3,775 posts

Awards Bar:

Posted 26 January 2017 - 04:17 PM

 

Do my transfers, and pray to the Gods

Indeed or I make offerings to the dwarves or both! I sing incantations too! hahahahaha  :cool: 


  • JustAnEyedea likes this

#38 JustAnEyedea

JustAnEyedea

    Destroy Your Television

  • Black VIP
  • 536 posts

Donator


Awards Bar:

Posted 26 January 2017 - 04:37 PM


Do my transfers, and pray to the Gods

Indeed or I make offerings to the dwarves or both! I sing incantations too! hahahahaha :cool:

For like 7 days straight until I see growth! Lolol

#39 Arathu

Arathu

    Dirtmaker

  • OG VIP
  • 3,775 posts

Awards Bar:

Posted 26 January 2017 - 04:53 PM

 

 

Do my transfers, and pray to the Gods

Indeed or I make offerings to the dwarves or both! I sing incantations too! hahahahaha :cool:

For like 7 days straight until I see growth! Lolol

 

Arathu's ............go to .......simple HWBRFA............... it's not a standard or anything like that,............it's just what I've found to work for me and the fungi.

 

40g powdered organic brown rice flower
9.5g powdered agar
500ml Hardwood tea

 

So what am I looking for?
 

  • I know the mycelium that we want (in this case) is going to be bright white in color.
  • I know that the mycelium we want (in this case) becomes rhizomorphic in its growth.
  • I deliberately use a known container/boundaries and further a known pattern (streaking) of inoculation within the boundaries in order to further chances of germinating, identifying, and then cultivating a specific species.

 

(Hardwood "cattative" can be anything you'd like to train a fungi to consume) at least that's my current view of the situation............that and every bit of the 7 days of rattles and smoke clouds................. hahahaha

 

 

A


  • CatsAndBats and JustAnEyedea like this

#40 JustAnEyedea

JustAnEyedea

    Destroy Your Television

  • Black VIP
  • 536 posts

Donator


Awards Bar:

Posted 26 January 2017 - 05:07 PM

Hardwood tea? Like boiled and strained? I've never come across that.
  • Arathu likes this





Also tagged with one or more of these keywords: artwork, friend

Like Mycotopia? Become a member today!