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c10h12n2o
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Quote:
Myco-pathic said: Here is the MS-GT plate #1 All t1’s cut from this plate. Still not a bacterial mess… Patch on the bottom right what the entire plate looked like At the start until the mycelium started to breach. That patch should breach soon. Just keeping it for study but it’s spent as far as my needs go.

The granular texture seen is LME that didn’t dissolve. I think the 10g/10g LME/MEA recipe was a bit heavy for 600 ml.
Adding Nutritional yeast for future plates. 10-MEA 2.5-YE 7.5LME @20 plates
That plate definitely IS a bacterial mess. This is very basic stuff.
Im not gonna keep repeating if you arent gonna hear it but clean myc is not slimy, those are not axenic cultures, there is definitely more than 1 organism present
I have no idea what you are talking about when you say "breach" but that plate is 100% definitely bacterial (at least)
This us what a clean culyure looks like 
Notice the leading edge.
This culture is not super rhiz , and a culture doesnt need to be rhizo to be clean, but even a tomentose culture should be fairly well organized, have a clean leading edge, and not be slimy
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  C10's Agar Guide + Tips and Tricks | c10's Flow Hood Build Guide "Partial knowledge is more triumphant than complete knowledge; it takes things to be simpler than they are, and so makes its theory more popular and convincing." "Convictions are more dangerous enemies of truth than lies" ― Friedrich Nietzsche
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Myco-pathic
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Re: Ms agar question [Re: c10h12n2o]
#27439996 - 08/23/21 06:48 PM (2 years, 8 months ago) |
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Malt Extract Agar is used for the cultivation of fungi and is not intended for use in the diagnosis of disease or other conditions in humans. An acidic medium which will support the growth of most yeasts and molds whilst inhibiting most bacteria.
(MEA)
Mycologists: are you referring to the light grey around the transfer wedge or the texture in the media?
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Myco-pathic
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Re: Ms agar question [Re: c10h12n2o]
#27440004 - 08/23/21 06:57 PM (2 years, 8 months ago) |
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Comment on the t1 .. you Quoted the ms plate and I already said it’s spent as I’m done with it.
Something is defiantly strange with the agar mix. It’s very very soft like it almost didn’t completely solidify.
That’s what I mean when I say the mycelium is growing though the top layer and not fully on top.
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Myco-pathic
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As close as I could get without the camera freaking out.
Again ignore the undissolved LME.


That soft ring will follow the growth out. Has a stranded texture. I have plates with bacteria and it’s way different. I really think this agar is too soft.
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shthd
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Quote:
Myco-pathic said: Here is the MS-GT plate #1 All t1’s cut from this plate. Still not a bacterial mess… Patch on the bottom right what the entire plate looked like At the start until the mycelium started to breach. That patch should breach soon. Just keeping it for study but it’s spent as far as my needs go.

The granular texture seen is LME that didn’t dissolve. I think the 10g/10g LME/MEA recipe was a bit heavy for 600 ml.
Adding Nutritional yeast for future plates. 10-MEA 2.5-YE 7.5LME @20 plates
OMG that lil guy running is so CUTE
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c10h12n2o
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lol... dude we know what MEA is. You said:
Quote:
Myco-pathic said: I think the 10g/10g LME/MEA recipe was a bit heavy for 600 ml.
Adding Nutritional yeast for future plates. 10-MEA 2.5-YE 7.5LME @20 plates
MEA = malt extract + agar + water (15g + 20g + 1L being pretty standard)
LME= light malt extract
MEA is the final product, agar powder and LME and water are the ingredients. this is why PBJ asked you if that was a typo. this could be what is "strange about the agar mix"
and you said:
Quote:
Myco-pathic said: Here is the MS-GT plate #1 All t1’s cut from this plate. Still not a bacterial mess… Patch on the bottom right what the entire plate looked like At the start until the mycelium started to breach. That patch should breach soon. Just keeping it for study but it’s spent as far as my needs go.

obviously i understood that you are done with that plate, the part i was taking issue with is where you said that it is "not a bacterial mess." it absolutely is a bacterial mess
the t1s do not look good, because you transferred them from bacterial colonies. might still fruit because bacteria =/= guaranteed failure, they usually fruit anyway. but the bacteria makes it much more susceptible to other contams (which might already be present as well)
as i said before, roll the dice on these if you want, but you should serial streak a fresh plate and start working toward an actually clean culture which you can use with 100% success whether these work out or notQuote:
Myco-pathic said: As close as I could get without the camera freaking out.
Again ignore the undissolved LME.


That soft ring will follow the growth out. Has a stranded texture. I have plates with bacteria and it’s way different. I really think this agar is too soft.
like we keep telling you, that does not look good. clean myc (tomentose or rhizo) is not slimy like that, there are multiple organisms present there
bacteria by itself does not look the same as bacteria mixed with cube myc. same with trich: trich myc by itself does not look the same as trich myc mixed with cube myc.
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  C10's Agar Guide + Tips and Tricks | c10's Flow Hood Build Guide "Partial knowledge is more triumphant than complete knowledge; it takes things to be simpler than they are, and so makes its theory more popular and convincing." "Convictions are more dangerous enemies of truth than lies" ― Friedrich Nietzsche
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frikiriki
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No offense, but i agree with most comments saying this is not good growth. You might be wasting your time on something that is not healthy/is stressed.
The spore syringes in general are a no from me. The spores cannot set and grow well in liquid.
I use sterile spore swabs and transfer part of the cotton swab to the plate. The cotton is clean and boosts growth time. You can tell a lot more quickly if a dish is good or not.
(you can use sterile spore swabs in conjunction with plates, just swipe and go)
(Also good for swabbing gills for spores and starting a library of your own. That way you don't have to purchase from vendors. )
Nice set up though...quite envious!
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Myco-pathic
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Re: Ms agar question [Re: frikiriki]
#27440062 - 08/23/21 07:42 PM (2 years, 8 months ago) |
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It was a print. Had syringes from way back. All of them went funky on me.
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Myco-pathic
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Re: Ms agar question [Re: c10h12n2o]
#27440078 - 08/23/21 07:51 PM (2 years, 8 months ago) |
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Your saying the agar mix is jacked… I was thinking something was up with it because before I bought the LME it was much more of what I expected.
I will scrape some more into some new mixes.
I really do appreciate everyone’s advice. Hang with me until I get it.
I have tons of material to keep trying with. It’s been 10 years since I fruited anything. So really looking forward to it!
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Myco-pathic
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Round 2 FIGHT!
Just using MEA solo this time.
Ordered some agar agar. Really disliking the naming scheme they used for agars
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c10h12n2o
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If you are using a premixed MEA, just follow the instructions on the packaging
Are you heating it in a media bottle on a hotplate? That might be part of your problem with dissolving, those arent made to be heated that way
Check out the guide in my sig
For the most consistent, clear, even agar, i like to first mix it into cold water, then boil it in a beaker in a microwave on low power and mix well
If you really want to use a hot plate, use a flask
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  C10's Agar Guide + Tips and Tricks | c10's Flow Hood Build Guide "Partial knowledge is more triumphant than complete knowledge; it takes things to be simpler than they are, and so makes its theory more popular and convincing." "Convictions are more dangerous enemies of truth than lies" ― Friedrich Nietzsche
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Myco-pathic
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Re: Ms agar question [Re: c10h12n2o]
#27440805 - 08/24/21 10:40 AM (2 years, 8 months ago) |
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No heat just stir bar being used.
I will check it out. New plates look much better. Very firm.
Last ones were like Jell-O shots. That’s what I meant by the mycelium submerged and breached. 😂
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The Mycologist
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Your agar mix is not to blame
-------------------- "That you are here—that life exists, and identity; That the powerful play goes on, and you will contribute a verse.” ― Walt Whitman, Leaves of Grass

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Myco-pathic
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Quote:
The Mycologist said: Your agar mix is not to blame
That comment was not very constructive or helpful.
Yes the agar was part of the problem as I stated above I remade the agar and I’m starting over and doing some things differently.
At the beginning this post I respectfully disagreed because not every comment on the Internet is the nugget of gold.
I haven’t been here in 10 years and I don’t know who knows what they’re actually talking about. Thankfully guys like C 10 are here to be credible and helpful. 👍🏻
The Shroomery has always been a friendly and engaging environment for a very nerdy hobby. Please don’t treat this like Reddit.👎🏻
Edited by Myco-pathic (08/24/21 12:10 PM)
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The Mycologist
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Bro, soft or hard agar. That growth isnt right.
You are blaming the rain for leaks in your roof.
-------------------- "That you are here—that life exists, and identity; That the powerful play goes on, and you will contribute a verse.” ― Walt Whitman, Leaves of Grass

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frikiriki
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They could just be shit genetics. (excuse my french)
Do you store them at room temp? Or fridge?
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PBJ710
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Quote:
The Mycologist said: Bro, soft or hard agar. That growth isnt right.
You are blaming the rain for leaks in your roof.

The agar firmness/nutrition content is something that needs to be addressed, but it's not the real issue on those plates.
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c10h12n2o
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Re: Ms agar question [Re: PBJ710]
#27441801 - 08/24/21 11:49 PM (2 years, 8 months ago) |
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Are you boiling your agar at all? You should really check out my agar guide in my sig
Like they said the agar isnt really the issue here. Thats more of a cosmetic thing.
Check this thread as well, it shows some examples of hidden contams
https://www.shroomery.org/forums/showflat.php/Number/22020260
Also, i just started a new project i will post in a few weeks (this thread and a few others gave me the idea). Basically i combined spores and clean myc with various contams to show what cultures look like when they are intermingled with various contams, should be pretty helpful for people.
Contams look different intermingled w cube myc vs on their own
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  C10's Agar Guide + Tips and Tricks | c10's Flow Hood Build Guide "Partial knowledge is more triumphant than complete knowledge; it takes things to be simpler than they are, and so makes its theory more popular and convincing." "Convictions are more dangerous enemies of truth than lies" ― Friedrich Nietzsche
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karri0n
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Quote:
Myco-pathic said:
I did use as a little liquid as possible I poor it very slowly until there is still gaps on the plate and then drop by drop until the liquid Connects.
  
This is legit nightmare fuel.
As I pictured your plate, pooling with bacterial slurry, I winced and shouted "NO!" with each and every drop, methodically and meticulously dripping away all hope from your gorgeous plates in front of your epic hood and immaculate lab environment.
You absolutely are cultivating bacteria and mold like this.
As little liquid as possible would be one drop. That would still be too much, and is likely not going to be axenic.
You should put a single drop onto an inoculation loop or sterile swab and streak several plates. This is the first thing you learn about cultivating mushrooms from spore on agar. Better yet, if you have a print, just streak the print itself using the swab or loop and skip the syringe altogether.
If you did this with the vendor syringes, that's also why you didn't have any success. I'd wager that you had better luck because you made this syringe in front of a hood, from a print,which had time to dry and wouldn't have motile bacteria suspended in water.
Water is basically the enemy on an agar plate, because it give bacteria the ability to be motile, whereas the 2d surface *should* give mycelium a motility advantage. If there is no water and 1 bacterial cell, the mycelium can outcompete it or destroy it with antibacterial metabolites. If there is water, that one bacterial cell can rapidly multiply and move anywhere else water is present, quickly out competing the mycelium.
If there are no bacteria at all, water isn't a problem(hence people do transfers of clean growth to plates with condensation/pooling and are ok sometimes), but there can't be truly axenic prints because mushrooms fruit in open air and necessarily will be exposed to bacteria and mold spores.
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Panaeolus Bisporus
Edited by karri0n (08/26/21 02:11 AM)
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shthd
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Re: Ms agar question [Re: karri0n]
#27443306 - 08/26/21 03:44 AM (2 years, 8 months ago) |
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I took their statement as referring to pouring their agar mix, not the spore solution. (They do mention they used a print after failed attempts with syringes.) That much spore solution would certainly lead to primordial soup lol
You say you haven't grown in 10 years? Where was the lab equipment stored in that time, specifically the flow hood? Is the HEPA filter new?
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