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Mushrooms, Mycology and Psychedelics > Mushroom Cultivation
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Re: AGAR ENVY! (Anything and All things agar!) [Re: huey.bluey]
    #28497851 -

huey.bluey said:
I want gentamicin for agar but its script only


You can buy vials meant for veterinary use without a script.
I’ve been tempted to buy some as well but it isn’t necessary moreso just may-possibly be beneficial in certain circumstances


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Re: AGAR ENVY! (Anything and All things agar!) [Re: MicroMycelium]
    #28497990 -

How would you guys transfer these? They're poured thin and one got chunky.






Pan Cambo Goliath and Pan Bisporus. Any advice would be appreciated.

Thanks.

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Re: AGAR ENVY! (Anything and All things agar!) [Re: NotFromGeorgia] * 1
    #28498004 -

Sure do nothing but mold as well.



Ghost germ plate and its transfers.





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“Sunny days wouldn't be special if it wasn't for rain.” – Curtis Jackson
“There is a world beyond ours, a world that is far away, nearby, and invisible.” – Maria Sabina

Edited by 0mar (10/09/23 08:18 PM)

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Re: AGAR ENVY! (Anything and All things agar!) [Re: 0mar]
    #28498049 -

Where’s the mold?
Those don’t look amazing but good enough to keep transferring until they do.
I’ve started to do T1 to water agar, you might want to give that a try.

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Re: AGAR ENVY! (Anything and All things agar!) [Re: 0mar]
    #28498074 -

that middle transfer plate looks weird. i'd take some from the first and last t1 plates though, the left colony on the last plate especially

that old germ plate looks like the same type of growth across it, and no mold spore colors. so i'm thinking if it is moldy, the cube myc is mostly winning

looking at them again, your new germ plates don't look terrible either, just new and weak. you used less nutes this time right? that'll make the growth thinner at first ime

i think you'll get something outta these but also :shrug:


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The most important step a man can take. It's not the first one, is it?
It's the next one. Always the next step, Dalinar.

Edited by NotFromGeorgia (10/09/23 09:12 PM)

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Re: AGAR ENVY! (Anything and All things agar!) [Re: mistermiyagi]
    #28498112 -

All the growth started out as clear, white dots appeared and then it gets white and really fluffy like stand off the plate fluffy. That’s why I think it’s mold it doesn’t look like any the myc I see here.


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“Sunny days wouldn't be special if it wasn't for rain.” – Curtis Jackson
“There is a world beyond ours, a world that is far away, nearby, and invisible.” – Maria Sabina

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Re: AGAR ENVY! (Anything and All things agar!) [Re: NotFromGeorgia]
    #28498122 -

I used less nutes but the growth looked the same regardless of recipe. All my growth starts out as the see through looking stuff and eventually turns white and puffy with strands sticking up. From my understanding myc starts off as white not see through.


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“Sunny days wouldn't be special if it wasn't for rain.” – Curtis Jackson
“There is a world beyond ours, a world that is far away, nearby, and invisible.” – Maria Sabina

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Re: AGAR ENVY! (Anything and All things agar!) [Re: 0mar]
    #28498202 -

Have you tried no pours? I did some agar work recently and for some reason I got growth that looked like that on my Petrie dishes but not on my no pours, however that looks bacterial or something. Sometimes they can be aerial like this


Yours looks different though at the same time... Did you cool your razor where the mycelium is supposed to be? I did that, but someone advised me to do it towards the edge so I can see the growth better, if you do it in the middle it makes it look funny like mine are. You can see where my razor was cooled.

This is what the no pours looked like.

Looks healthier... Same recipe though, only difference was the containers. 🤷

Honestly those plates probably contaminated during your transfer procedure and the mold won, or whatever it is. That happened to me when I tried to take clones the first time.

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Re: AGAR ENVY! (Anything and All things agar!) [Re: 0mar]
    #28498206 -



Spent most of yesterday making corn meal agar and autoclaving it to pour. Tonight I got to plate some spores.

Never did any of this before but it was really fun.

Included some peptone and ampicillin just to up my success rate.

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Re: AGAR ENVY! (Anything and All things agar!) [Re: Subhuti] * 2
    #28498252 -

Peptone is good food for bacteria. Ampicillin is unnecessary as transfers and clean media alone are groovy enough to generate axenic cultures. Using antibiotics for basic culture work to make up for possible sloppiness on behalf of the cultivator is unwise and it would be better to see when you mess up anyway. something like ampicillin could only inhibit visible growth without killing the bacteria so when it goes to grain the bacteria is no longer inhibited and can flourish.

Malt, a pinch of yeast, agar, and water is the best set of ingredients I've found so far for most fungi.

Very much respect to you for taking the dive into agar work. Don't let me sway you from using those dishes either. They'll likely work just fine.

That is a lovely lab bench and some nice looking glassware you are working with there.


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Question everything, it's very responsive.

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Re: AGAR ENVY! (Anything and All things agar!) [Re: Nichrome] * 1
    #28498257 -

Thanks this is all great to know. It wasn’t my idea to use the ampicillin but the person teaching me how she makes agar plates and basic lab sterilization practices felt very strongly that including the ampicillin couldn’t hurt.

In the future I’ll exclude them. I think we did a solid job keeping clean though. Autoclave for medium, then Bunsen burner and ether to flame sterilize the glass and create a sterile field while we poured. Then half an hour in the sterile field on the bench before flipping them and putting them into a sterile incubator overnight to dry a bit.

Edit: in the future I’d like to use malt extract instead of corn meal. It just wasn’t an option this time.

Edited by Subhuti (10/09/23 11:49 PM)

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Re: AGAR ENVY! (Anything and All things agar!) [Re: Subhuti]
    #28498273 -

Subhuti said:
Thanks this is all great to know. It wasn’t my idea to use the ampicillin but the person teaching me how she makes agar plates and basic lab sterilization practices felt very strongly that including the ampicillin couldn’t hurt.

In the future I’ll exclude it. I think we did a solid job keeping clean though. Autoclave for medium, then Bunsen burner and ether to flame sterilize the glass and create a sterile field while we poured. Then half an hour in the sterile field on the bench before flipping them and putting them into a sterile incubator overnight to dry a bit.


This bit is still common practice in standard college bio teachings but is very outdated and you may find does not work very well for mycology. There are a lot of vectors for contamination to occur in that process. Look into the term "SAB" or "still air box" or more fitting for your fancy lab would be a nuclear grade laminar flow hepa filter with a horizontal flow, aka "flow hood" or "fan filter unit". These are the gold standard in biology labs worldwide. 

I thought you probably did that bunson/incubator method from the look of your lab and the fact that your dishes are not wrapped with film and are sitting out in the open. Using the bunson method typically requires the use of strong antibiotics and selective biocides in the media. That method was used before many modern practices were developed.

Keep coming back. This place albeit flawed like any place, is truthfully the best knowledge base for modern practical mycology methodology. All the best innovation and discovery happens in garages and basements with makeshift devices used by people of madness. The Tesla's and the Wright's of the world. All the labs and fancy lab coats get to work with sloppy seconds and are just riding the coat tails of the real hero's out there putting madness to practice in a practical fashion.

Personally I'd watch those unwrapped plates for like 5-7 days before trusting them for any usage that is of value to you.


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Question everything, it's very responsive.

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Re: AGAR ENVY! (Anything and All things agar!) [Re: Nichrome]
    #28498282 -

This is great to know because I’m just basing what we did off of my partner’s non-mycology background. I have a SAB at home.

In the future I can sterilize the agar and then would it be wise to pour and wrap with parafilm? Or is it a different sort of film that you’d use?

I could wrap them in film and drop liquid culture on them in the SAB and then rewrap them?

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Re: AGAR ENVY! (Anything and All things agar!) [Re: Subhuti] * 1
    #28498293 -

Parafilm and clingwrap are the two commonly used films.

A lot of new people use clingwrap but imho it has many, many, many downfalls and as a lab material is much less useful than parafilm which has many uses. Parafilm is harder to learn how to use correctly and I see few people here doing so hence the draw to clingwrap. You'll even see "team clingwrap" in peoples signatures indicative of some kind of "loyalty" which is a mental trip centered on social acceptance and enabling.

I love parafilm as a lab stuffs but it does work especially well for wrapping plates. It does not draw spores and dust from the environment with static the way cling does, and it has a wax coating which traps up any dust or spores that do land on there rendering them immobile and all stuck up in sticky shit.

Yes, pour hot in the SAB or in front of the flow, and wrap with film once cool. Keep them in zipper bags to keep dust off them.


--------------------
Question everything, it's very responsive.

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Re: AGAR ENVY! (Anything and All things agar!) [Re: Nichrome]
    #28498300 -

Thank you very much for all these ways to improve. I took notes for next time.

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Re: AGAR ENVY! (Anything and All things agar!) [Re: Subhuti] * 2
    #28498308 -

A big ass sterilite tub I melted two arm holes in with a piping hot coffee can being enough to make anexic cultures in blows my mind, but it works. Second vote of confidence for malt, agar, yeast, and water. It’s a great medium.


--------------------
A Love Letter to New Growers
The 7 Edicts of Mushroom Cultivation as Decreed by Smellyhobbit

:kodama:


Anyone can get a canopy. But can you get what you want?

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Re: AGAR ENVY! (Anything and All things agar!) [Re: Smellyhobbit]
    #28498318 -

On the note of the clingwrap/parafilm argument, I tried using my old parafilm, and the paper doesn't stay on any more and that made me nervous so I stopped using it. But I found my old cut roll of clingwrap, and even though I remember hating it, it may be better than putting them back into the ziplocks they came out of.


--------------------
"Remember, kids, the difference between science and screwing around is writing it down" -adam savage
Flowchart for Recommended plan of action.
Learn the tried and true way to grow mushrooms
Use the Damn search engine
After you know what you're doing, take a break 
Pick a book, Make some chips!
Josex said:Don't take the site seriously bro, ain't worth it.
 

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Re: AGAR ENVY! (Anything and All things agar!) [Re: Subhuti]
    #28498334 -

Just fyi it's been said before that unless you need antibiotic agar to clean up really dirty stuff, it just makes it so you can't see the contams to transfer away from thereby making your efforts worthless, because as soon as you take the agar to grain the baddies will show back up, if I'm not mistaken by what I read. Just saying... Water agar would be a better option. And learning proper sterile technique. It's like putting a scope on a pistol... Just learn to shoot.

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Re: AGAR ENVY! (Anything and All things agar!) [Re: Nichrome]
    #28498336 -

I don't see the issue, they both work. Dust cannot pass through cling wrap so I don't see your point, at all. Guess what, all that shit will stick to wherever the parafilm isn't so.... Team whatever works for you, because they both work perfectly. If your shit contams, you fucked up period.

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Re: AGAR ENVY! (Anything and All things agar!) [Re: TrKyNcKmSk]
    #28498356 -

I wasn't really making a point I guess, more just making conversation.
The clingwrap protects itself, the parafilm needs that paper layer to keep the underside clean. If I can't cut strips without compromising the parafilm, then it's kind of useless, no?


--------------------
"Remember, kids, the difference between science and screwing around is writing it down" -adam savage
Flowchart for Recommended plan of action.
Learn the tried and true way to grow mushrooms
Use the Damn search engine
After you know what you're doing, take a break 
Pick a book, Make some chips!
Josex said:Don't take the site seriously bro, ain't worth it.
 

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Mushrooms, Mycology and Psychedelics > Mushroom Cultivation
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