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Agar experimenting


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#1 Microbe

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Posted 29 October 2014 - 10:40 AM

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While conducting g2g i spilled some grain and of course some went on the floor. I decided this would be perfect oppurtunity to set myself up for some practice at isolating away from a contamination.

What i did.....

Opened to agar plates in front of flow hood and picked up some colonized grain off the floor that has been stepped on and had lint stuck to it. I placed a single piece in each plate and placed into incubation.

After doing this i said to my self, "self, WTF were you thinking" and placed into incubation. Over the next several days i watched as the bacterial contamination showed up around the grain. On the 7th day i noticed some mycelium poking out under the grain so i tilted the jar to roll the grain to the edge and the above picture is what i captured.

What im wanting to observe is just exactly what this mycelium does. Im certain its going to retreat but if it consumes the bacteria then i would think that as long as its a good fruiter then maybe i should isolate it yes? When i first started agar work a plate like this always ended up in the decon bucket and never produced any viable myc. I would love to have a microscope to possibly identify this bacteria! I was scammed, well not scammed because i received my money back, but i found one that was listed for 1299 on clearance for 749 and waited almost six months to receive im sorry were unable to fullfill your order! Very disappointing! !!

Edited by coorsmikey, 26 May 2016 - 06:36 PM.
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#2 Arathu

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Posted 29 October 2014 - 06:50 PM

You will fill up many notebooks and pitri plates in pursuit of those rhizomes........and what they become. Isolate those that strike you as interesting and exhibit characteristics you are looking for. I like to test substrates and transfer mycelium from agar to wood directly. Agar work is fun............  :meditate: 


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#3 Microbe

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Posted 30 October 2014 - 02:02 AM

e61e9886097964315f5cd0276f772efd.jpg

While conducting g2g i spilled some grain and of course some went on the floor. I decided this would be perfect oppurtunity to set myself up for some practice at isolating away from a contamination.

What i did.....

Opened to agar plates in front of flow hood and picked up some colonized grain off the floor that has been stepped on and had lint stuck to it. I placed a single piece in each plate and placed into incubation.

After doing this i said to my self, "self, WTF were you thinking" and placed into incubation. Over the next several days i watched as the bacterial contamination showed up around the grain. On the 7th day i noticed some mycelium poking out under the grain so i tilted the jar to roll the grain to the edge and the above picture is what i captured.

What im wanting to observe is just exactly what this mycelium does. Im certain its going to retreat but if it consumes the bacteria then i would think that as long as its a good fruiter then maybe i should isolate it yes? When i first started agar work a plate like this always ended up in the decon bucket and never produced any viable myc. I would love to have a microscope to possibly identify this bacteria! I was scammed, well not scammed because i received my money back, but i found one that was listed for 1299 on clearance for 749 and waited almost six months to receive im sorry were unable to fullfill your order! Very disappointing! !!

I have many notebooks, and since day 1 i have only loved the one the one that i loved.....

#4 torn2bits

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Posted 30 October 2014 - 04:30 AM

. Makes sense, that if Myceleum caulk overcome common bacteria that it would be a hearty fruiting strain.
I like your theory that if given time that myceleum will isolate itself on agar.
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#5 Microbe

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Posted 30 October 2014 - 03:42 PM

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I think the mycelium might actually colonize this plate. Its hard to tell in the pic probably, my phone cam wants to focus on the ziploc bag instead of the agar surface but it appears that it is advancing over the bacteria.

Edited by Microbe77, 30 October 2014 - 03:44 PM.

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#6 Arathu

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Posted 30 October 2014 - 04:47 PM

If that lower left half isn't enticing I don't know what is. Personally my first move would take that 1/2 in a first transfer. It looks like nice rhizome will be available shortly from that..................my 2c...........I don't like to mess with bacteria too much, fungi is another thing all together.

Actually a dozen, even two dozen new plates could be made right now.......hmmmm.....I guess depends on how you want to play it.  :cool:


Edited by Arathu, 30 October 2014 - 04:52 PM.

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#7 Microbe

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Posted 01 November 2014 - 08:35 AM

Im about to transfer it. I have seen enough on this plate and am certain that this myc will compete well with a bacteria, at least this type of bacteria, and i think i will get some rhizo after a few transfers. There is a lot going on in this plate and there some more myc competing with some bacteria on the right side.

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#8 Microbe

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Posted 01 November 2014 - 08:41 AM

I want to transfer from the bottom edge that is advancing over the bacteria but have some questions.
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Do i use a inoculation loop and transfer via smear in an attempt to leave the bacteria behind? Ot do i transfer a agar plug to bring the bacteria with so that it can continue to compete and develop with the presence of thos bacteria? Maybe i can do both....

Edited by Microbe77, 01 November 2014 - 09:15 AM.

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#9 Microbe

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Posted 01 November 2014 - 08:43 AM

And im not sure this is Texan but just called it that after hearing that mentioned along with PE6. A few months ago i was working with what supposed to be PE bit turned out not to be or maybe the mutation reversed itself when i germinated on agar and made several transfers i dont know.

Edited by Microbe77, 01 November 2014 - 08:44 AM.


#10 torn2bits

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Posted 01 November 2014 - 10:35 AM

Rumor has it that PE is unstable, so many people attempted isolating & ended up with a non fruiting strain.
Micro, I still think you should be named AgarWizzard.
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#11 Microbe

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Posted 01 November 2014 - 10:54 AM

Rumor has it that PE is unstable, so many people attempted isolating & ended up with a non fruiting strain.
Micro, I still think you should be named AgarWizzard.

Maybe i should have went spore solution>grain>bulk sub>fruit then clone..... oh well i gave my syringe away, maybe i will order another sometime but right now im working with 3 strains at once right now and its keeping me busy.

#12 Dlight

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Posted 01 November 2014 - 11:07 AM

  What are  rhizome?  What are incubation conditions for dishes? I have a glove box not a flowhood.  How would I do it differently than you have?  I want to get into agar experiments.  I think the supplies are on the way.  



#13 Microbe

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Posted 01 November 2014 - 11:07 AM

Microbe77 nice work . I'm not advanced enough to work with agar, I look forward to watching your work.

Thank you for the kudos but i dont think it takes advanced skill set to work with agar. It was the first thing i worked with when i dove head first into this hobby. It started off just germinating spores and growing out inoculate. Im always saying that agar work is the backbone of the hobby and it has been very rewarding. It takes patiences and alot of luck or firing at the hip, especialy when working without i microscope, but the rest will come from experience rather failures or success.
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#14 Microbe

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Posted 01 November 2014 - 11:29 AM

What are rhizome? What are incubation conditions for dishes? I have a glove box not a flowhood. How would I do it differently than you have? I want to get into agar experiments. I think the supplies are on the way.

Rhizomes are tubular like structures like a root or roots of a plant. Im not certain but i think rhizomes in mycology is a reference to the Hyphae structure. Rhizomorphic growth looks almost like roots branching out across the surface. This typicaly what we look for and it is probable that it will be agressive colonizer, fruiter, or both.

I incubate at 75 F but they can be stored anywhere you keep your spawn jars including room temperature. I even apply the brakes on my plates at times by tossing them in the fridge for a few weeks or even a few months. Im still trying to determine where the cutoff is for cold storage of the colonized plates when a slant is required. Anyway back to the topic, glove box is fine and just follow sterile technique as usual for a glovebox. I started in a glove box, and will be working in one with this contaminated plate so my flowhood doesnt blow this crap all over the place. But my first spore to agar germination was in a GB. Something i did was.install lights on my glove box because you need alot of light when you start selecting specimens to transfer. Any question from making nutrient agar to isolating let me know and i will do my best to answer. Im love it when i see people getting ready to start agar work. I gaurantee everyone at some point thought man alive i didnt i do this sooner.

Edited by Microbe77, 01 November 2014 - 11:34 AM.


#15 Microbe

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Posted 01 November 2014 - 11:46 AM

Here is a no pour method i use. And i also includes a pic of my glove box to show you how i illuminate my working area. I would turn all the lights in the room and man alive the mycelium was bright!

https://mycotopia.ne...Agar-Jars......

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#16 Heirloom

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Posted 01 November 2014 - 01:27 PM

we are all drawn to different things and still our paths cross , teaching.

that glovebox screams dedication.

I got few few pics of maz on rye that stayed in jar.



good luck finding a microscope, like to get one myself.

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#17 Microbe

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Posted 01 November 2014 - 04:22 PM

Im not sure why i call it a Glove Box because its a Still Air Box but i still find myself calling a glove box. Break the seal in agar and you will be rewarded.
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#18 Microbe

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Posted 01 November 2014 - 04:22 PM

we are all drawn to different things and still our paths cross , teaching.

that glovebox screams dedication.

I got few few pics of maz on rye that stayed in jar.



good luck finding a microscope, like to get one myself.

You should see my flowhood! Lol

#19 Microbe

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Posted 01 November 2014 - 04:25 PM

. Makes sense, that if Myceleum caulk overcome common bacteria that it would be a hearty fruiting strain.
I like your theory that if given time that myceleum will isolate itself on agar.

My theory on isolation is that if the mycelium is given enough surface area it will isolate itself without any intervention. Im about to document my agar work for you ToRn.

#20 hyphaenation

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Posted 01 November 2014 - 08:04 PM

Yah looks like search is broken again ...

 

For a long time i've just gone to google and typed mycotopia search-term.

 

Like:

 

mycotopia flowhood

mycotopia glovebox

mycotopia waylitjim woodlovers

mycotopia buckaroo banzai popcorn slurry

 

etc.

 

I find it works quite well


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