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Mushrooms, Mycology and Psychedelics > Mushroom Cultivation
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Re: AGAR ENVY! (Anything and All things agar!) [Re: D3_Myc]
    #26980683 -

I got a case of 229694 celltreat plates from amazon a couple months ago. They’re a lot stronger than the Thomas 7453 I was using before. The lids don’t fit as snugly, the edges are rounded, and the sleeve doesn’t come with the slit in the side to make it easier to open but other than that I like them. They come in sets of 20 as opposed to 25 so I can actually unload them vertically in my sab :kenthumbup:


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Re: AGAR ENVY! (Anything and All things agar!) [Re: Yeetusdeetus]
    #26980727 -

Big shoutout to the Shroomery and Myers Mushrooms for teaching me how to do agar.

Got my flowhood unboxed today and poured plates same day! Boom!


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Re: AGAR ENVY! (Anything and All things agar!) [Re: nogoodnamesleft]
    #26980736 -

Gorgeous flowhood!

Also really enjoying D3monics posts! <3


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:computertrip:

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Re: AGAR ENVY! (Anything and All things agar!) [Re: GrinchGrower]
    #26981069 -

Well I ended up shaking my jars that I had just inoculated, it just started on a cluster of grains. I would say it was only about 5% totally inoculated. But I went by what you said and to shake at that time. I thought you were supposed to wait until about 20-30 percent colonization and then shake. I felt like maybe I should of waited because following by what you said I feel I shook it up too early, which in return won’t be as effective when you do it too early correct? Shaking  early and too many times isn’t good right? I feel like things are messed up now instead of waiting until the 20 - 30 percent to shake, and I did it early that I feel like I e definitely messed something up now...


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Edited by psycho_nauticus (10/12/20 10:48 AM)

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Re: AGAR ENVY! (Anything and All things agar!) [Re: psycho_nauticus]
    #26981394 -

psycho_nauticus said:
Well I ended up shaking my jars that I had just inoculated, it just started on a cluster of grains. I would say it was only about 5% totally inoculated. But I went by what you said and to shake at that time. I thought you were supposed to wait until about 20-30 percent colonization and then shake. I felt like maybe I should of waited because following by what you said I feel I shook it up too early, which in return won’t be as effective when you do it too early correct? Shaking  early and too many times isn’t good right? I feel like things are messed up now instead of waiting until the 20 - 30 percent to shake, and I did it early that I feel like I e definitely messed something up now...


Do what makes you comfortable. Most will recommend to shake at 30% and shake again at 100%. After the myc recovers you're good to spawn. Its proven. It works. Most are also using big wedges. I used small wedges and it worked fine. I believe the idea behind multiple shakes from a small transfer is to get the myc distributed throughout the jar instead of waiting a long ass time to get the jar to full colonization from such a small area of growth and risking the possibility of the grain drying out.


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Edited by Doctor Mario (10/12/20 12:38 PM)

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Re: AGAR ENVY! (Anything and All things agar!) [Re: Doctor Mario]
    #26981440 -

I think lotkids method is sound. Now that you’ve shaken it I would wait til it hits 30% and shake again.

The way he does it just makes 10x the inoculation points at the expense of setting the myc back like a day. Should definitely be faster overall.


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LAGM2020

Edited by A.k.a (10/12/20 02:30 PM)

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Re: AGAR ENVY! (Anything and All things agar!) [Re: A.k.a]
    #26981571 -

How long has it been? Keep an eye on recovery time, otherwise it should be fine.


--------------------
"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: A.k.a]
    #26981589 -

I'm currently running an experiment that to my knowledge is novel in shroomery history - if it has been done before it would be great to see a link.

It is a technique I found in this Chinese research paper:

"A handy method to remove bacterial contamination from fungal cultures"

https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0224635

They used this method to free fungal cultures from bacteria. In their case they did it with molds, which they intend to grow for reasons beyond the understanding of people like us, people who grow real mushrooms :laugh:

The idea is that they observed that mycelium can grow through and within agar, while bacteria can only grow on its surface.


So what they did is:

Take a fresh agar plate.
Cut a hole in the middle.
Place a piece of contaminated agar in it.
Then place a sterile microscopy cover slide over it, make sure it lays perfectly flat.


They found out with great success, that the mycelium finds its way out of this trap while the bacteria stays behind. It is all in the paper, with photos and all.


This idea is in line with the sandwich tek like here (scroll down or better download pdf):
https://www.researchgate.net/publication/248894057_A_simple_and_effective_method_for_the_elimination_of_bacteria_from_fungal_cultures#read ;

It also shares the idea with this tek (the full article is behind a paywall, but the abstract says what the core tek is):
https://onlinelibrary.wiley.com/doi/abs/10.1046/j.1439-0434.2001.00662.x

Also the hot pour that has been known in mycology since long times is a similar idea.

What I like about the cover-slide tek  -  if it works  -  is the simplicity and ease of doing it.

What I do not yet know is, if it works with real mushroom mycelium. It will have some fighting to do to travel about 10mm though agar without oxygen - so I am not sure if it will work.

The chinese researchers also found out that with the molds they tried it with, the final clean culture was a very well defined and nicely growing mold - better than the original parent plate. They think it has to do with only the strongest mycelium making it through to the edge of the cover slide.

If it works, it would be a great addition to mycology agar teks. Hopefully leading to a faster way of getting clean cultures from dirty prints, clones, etc.

This is my experiment, pan peace river from the dirtiest spore plate I could find under this cover slide:


I will update when I have good or bad news. The papers I linked above are quite interesting to read for agar-enthusiasts.

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Re: AGAR ENVY! (Anything and All things agar!) [Re: xspak]
    #26981644 -

xspak said:
I'm currently running an experiment that to my knowledge is novel in shroomery history - if it has been done before it would be great to see a link.

...

I will update when I have good or bad news. The papers I linked above are quite interesting to read for agar-enthusiasts.


cool. reading. waiting for sushi door dash. ordering glass.

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Re: AGAR ENVY! (Anything and All things agar!) [Re: nmd_myco]
    #26981663 -

Sounds like the sandwich or hot pour techniques


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LAGM2020

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Re: AGAR ENVY! (Anything and All things agar!) [Re: GrinchGrower]
    #26981684 -

So you shake a total of 3 times? I thought you were only supposed to shake one time at 20 to 30 percent colonization, and then one more time when spawning. If you shake it too early like you did then doesn't that mess with the growth and strength of the mycelium regaining itself and starts to become less effective?


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Re: AGAR ENVY! (Anything and All things agar!) [Re: psycho_nauticus] * 1
    #26981688 -

You can expand myc nearly infinitely. Itll be fine


--------------------
"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: tryptkaloids]
    #26981710 -

No-pour day! Filled up a shoebox with these, we'll see how long that lasts me.





And here's a T1 of a TOC clone I took from a fruit that was showing some PE traits (fat knobby stem, small cap, light spore-load.)



I dropped a wedge onto grains, we'll see how it goes. The cloned fruit was on the small side, so hopefully there'll be some size variation.


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:sporedrop:  redbee's LAGM 2021  :sporedrop:

Edited by redbee (10/12/20 03:07 PM)

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Re: AGAR ENVY! (Anything and All things agar!) [Re: A.k.a]
    #26981786 -

So do you also do three shakes instead of two? Do you shake it first right when it becomes inoculated? And then AT 30 percent and then 100 percent? Or only do your first shake at 30 percent?

This is my 3rd attempt at RW, and I really don't want to fuck anything up again this time by not following correct procedure...


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Edited by psycho_nauticus (10/12/20 04:30 PM)

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Re: AGAR ENVY! (Anything and All things agar!) [Re: psycho_nauticus]
    #26981804 -

I'm very hands off and often only shake once because I try to avoid hovering so before I look at em they're at like 50%


--------------------
"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: tryptkaloids]
    #26981817 -

I know, but too late now since I already did. I know you're supposed to wait until getting to about 20 - 30 percent, which I've aways done for my jars. I was thinking of it giving it a try, but all today I've been thinking back on it wishing I didn't do it because I've heard countless times to not shake it too early and this disrupts the mycelium more, especially if having to shake it a second time and then a third time at spawning, with each time using more energy to recover. I wish I hadn't... :frown: I hope it doesn't mess up my jars or colonization or anything like that because then it will be my fault and fuck up again by following improper advice.

And since this is my 3rd time with RW everything was going perfectly until I decided to make that irresponsible decision...


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Re: AGAR ENVY! (Anything and All things agar!) [Re: psycho_nauticus]
    #26981902 -

Another method outside sandwiching is water agar. Just agar and water, myc rapidly spreads searching for food while bacteria is stalled from starvation. Misty has recommended it with success. I haven’t had to try it personally but I keep meaning to pour a short stack of them to keep on hand.


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D3’s Glog
D3's Agar TEK
Agar Porn Gallery
RW x MelMak Grow Along Thread

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Re: AGAR ENVY! (Anything and All things agar!) [Re: psycho_nauticus]
    #26981907 -

The early shake was shared with us by a tc who runs massive amounts of spawn.

You're stressing too much.

If your spawn is clean it will be fine, otherwise a shake had nothing to do with failure.


--------------------
"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: A.k.a]
    #26981990 -

A.k.a said:
Sounds like the sandwich or hot pour techniques


Yes, they are related.

The idea is the same for all - that bacteria can only live on the surface but not penetrate the agar.

So far I am aware of these teks:

- hot pour

- sandwich

- bottom pick (meaning you flip the agar and take a piece from the bottom of it, careful not to go too deep to get bacteria from the surface)

- Raper's ring tek - that is a tek where you heat a glass ring (a  'Raper's ring' - after the scientist Raper who invented the tek in 1937) so that it melts somewhat into the agar and place that hot ring in a plate then then your piece of contaminated agar in the middle - then the mycelium has to crawl under it, leaving the bacteria behind

- the partial-agar tek:  meaning you pour a plate so that only part of the dish is covered and then let the mycelium grow over it then the mycelium will try to crawl onto the empty part of the dish in search for food - leaving the bacteria behind - and there you can be able to take a clean piece of it

It is always good to know many teks to pick the one that is most suitable for the particular circumstance. The cover slide tek by these chinese guys may be nice for example for people who already work with microscopy and have slides around.

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Re: AGAR ENVY! (Anything and All things agar!) [Re: D3_Myc]
    #26981997 -

D3monic said:
Another method outside sandwiching is water agar. Just agar and water, myc rapidly spreads searching for food while bacteria is stalled from starvation. Misty has recommended it with success. I haven’t had to try it personally but I keep meaning to pour a short stack of them to keep on hand.


Thank you a lot D3monic ! That sounds like something that I imagine to work well. And not too hard to prepare along the side when one does an agar run anyway.

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Mushrooms, Mycology and Psychedelics > Mushroom Cultivation
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