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Rumpleforeskin
Scientist new to mycology



Registered: 07/05/20
Posts: 82
Loc: Canada
Last seen: 3 years, 1 month
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Quote:
Pastywhyte said: I usually go for 20 most of the time and 45 for grain water. Never really had issues but I don't work with yeasts or peptone. Usually just work with whats laying around the house 
I have had grain water LC go bacterial if I don't run a full 45 min.
Can I get some clarification? 20g AA to 1 L of grain soak water? TIA
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vsdlmao
Stranger


Registered: 10/03/20
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Re: BOD's Comprehensive Agar TEK [Re: bodhisatta]
#26968543 - 10/04/20 04:39 AM (3 years, 3 months ago) |
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Is there any point in using distilled water to get rid of the minerals added by tap water, or is the mineral content negligible enough so that it doesn't really make a difference?
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SubjectX
Stranger
Registered: 10/11/20
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Re: pour agar TEK [Re: vsdlmao]
#26983361 - 10/13/20 01:09 PM (3 years, 3 months ago) |
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OMG, this is one of the greatest explanations of anything I've ever seen. I know the thread is a little old, but I just had to say BRAVO!!. I think I can do this.
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verum subsequentis
seeker of truth



Registered: 03/22/16
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Re: pour agar TEK [Re: SubjectX]
#26983367 - 10/13/20 01:14 PM (3 years, 3 months ago) |
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You can absolutely do this.
This is one of those threads that doesn't get old. People will always be trying to learn agar and threads like this are here exactly for that reason.
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EvilSpawnJar
Bad Batch


Registered: 11/30/17
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Re: BOD's Comprehensive Agar TEK [Re: bodhisatta]
#26984957 - 10/14/20 11:38 AM (3 years, 3 months ago) |
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One question regarding agar color. How to get such a nice clear red agar as seen in first post?
I used some food colors before, but i get very "muddy" colors. I only tried agar with LME so far. And i assume LME makes it "muddy" looking. So any color i would add will be affected by "base" color of LME agar.
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verum subsequentis
seeker of truth



Registered: 03/22/16
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Re: BOD's Comprehensive Agar TEK [Re: EvilSpawnJar]
#26984966 - 10/14/20 11:41 AM (3 years, 3 months ago) |
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i like blue. I use about four or five drops per liter of agar. Red is fine but just know, It's made from a bug that has anti fungal properties. Use too much and you myc won't like it at all.
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EvilSpawnJar
Bad Batch


Registered: 11/30/17
Posts: 31
Last seen: 3 years, 2 months
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I asked about red, because that's what is posted. No need to be red. I have blue, yellow and red food colors.
But i don't know which agar recipe would be best to get nice and clear color. Like i said, when using LME, "base" color is deep brown, so any color is affected by it. And it will never be clear / see through, as is visible on 3rd image (melted agar)
Well color is not the most important thing. But it looks great.  For example, when i added blue, it ended up as dark turquoise or something like that.
Edit: I just noticed, you have mountain of plates in your sig. Nice stack. I'm just starting to get back into it again.
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Edited by EvilSpawnJar (10/14/20 12:00 PM)
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Brian Jones
Club 27



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Re: BOD's Comprehensive Agar TEK [Re: vsdlmao]
#26985005 - 10/14/20 12:07 PM (3 years, 3 months ago) |
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Quote:
vsdlmao said: Is there any point in using distilled water to get rid of the minerals added by tap water, or is the mineral content negligible enough so that it doesn't really make a difference?
There is no need for distilled water, unless maybe you live some place with awful water. The mineral content is either negligible or not undesirable. I think nearly everyone uses tap water.
-------------------- "The Rolling Stones will break up over Brian Jones' dead body" John Lennon I don't want no commies in my car. No Christians either. The worst thing about corruption is that it works so well,
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footpath
ὕδωρχοίρος

Registered: 07/16/19
Posts: 1,367
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Re: BOD's Comprehensive Agar TEK [Re: EvilSpawnJar]
#26985013 - 10/14/20 12:11 PM (3 years, 3 months ago) |
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Quote:
EvilSpawnJar said: I asked about red, because that's what is posted. No need to be red. I have blue, yellow and red food colors.
But i don't know which agar recipe would be best to get nice and clear color. Like i said, when using LME, "base" color is deep brown, so any color is affected by it. And it will never be clear / see through, as is visible on 3rd image (melted agar)
Well color is not the most important thing. But it looks great.  For example, when i added blue, it ended up as dark turquoise or something like that.
Edit: I just noticed, you have mountain of plates in your sig. Nice stack. I'm just starting to get back into it again.
It sounds like you're using too much of whatever you're using (dye or LME) if it's getting 'muddy' (opaque?).
This is a backlit plate made with the standard 10g agar agar/10g LME:

There's not really much need to be any more translucent than that. Nor would I consider it 'muddy'.
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EvilSpawnJar
Bad Batch


Registered: 11/30/17
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Re: BOD's Comprehensive Agar TEK [Re: EvilSpawnJar]
#26985036 - 10/14/20 12:21 PM (3 years, 3 months ago) |
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Just like you said. I use 10g LME and 10g A-A for 500ml. But i also add 1g yeast. Found it in Frank's agar media journal.
No need for yeast? and is it yeast that could make it "muddy"? Might post some pics,.. just need to take some.
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footpath
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Re: BOD's Comprehensive Agar TEK [Re: EvilSpawnJar]
#26985040 - 10/14/20 12:23 PM (3 years, 3 months ago) |
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Couldn't say - I haven't used yeast for culturing media. However, when I make bread, it sure does muck up that water.
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EvilSpawnJar
Bad Batch


Registered: 11/30/17
Posts: 31
Last seen: 3 years, 2 months
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Re: BOD's Comprehensive Agar TEK [Re: footpath]
#26985055 - 10/14/20 12:34 PM (3 years, 3 months ago) |
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Quote:
footpath said: Couldn't say - I haven't used yeast for culturing media. However, when I make bread, it sure does muck up that water.
Very good point. So i guess i will try next agar without yeast. Will have to do it soon anyway.
Does anyone use smaller plates as well, or only 100mm? I have 100mm, but i also bought 60mm. I was thinking that since they are smaller, you can just throw one full "plate" in jar. No need to split it in case of regular jars (not wide mouth).
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Edited by EvilSpawnJar (10/14/20 12:36 PM)
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footpath
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Re: BOD's Comprehensive Agar TEK [Re: EvilSpawnJar]
#26985082 - 10/14/20 12:47 PM (3 years, 3 months ago) |
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People sure do. However, even with 60mm plates, you'd be wasting a lot of potential by dropping the full agar puck into a quart jar. Seeing as people expand those little no-pour PP5 sauce cups (about the size of a US nickel at the base) to quarts, it'd be a better use of the culture to split it up.
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EvilSpawnJar
Bad Batch


Registered: 11/30/17
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Re: BOD's Comprehensive Agar TEK [Re: EvilSpawnJar]
#26985099 - 10/14/20 12:57 PM (3 years, 3 months ago) |
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Ok. Thanks for info. 
I thought, since you cut it, it will need to "recover". But i guess that's not really any significant time lost.
I guess i have too much of a weed growing mindset, cutting clones etc.
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Kontiki
Stranger

Registered: 09/12/19
Posts: 15
Last seen: 10 months, 25 days
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Hey guys, I just made my first Agar a couple of days ago and it went quite well  After cooling a bit I took some tissue pieces from a B aspiring to get some nice clones, could anyone give me some help/how to think now when I’m about to make the first isolation. Which plate/ which part do I choose for the first transfer?

Cheers all!
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funky123
Pilzfreund


Registered: 08/27/17
Posts: 257
Loc: Austria
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Re: pour agar TEK [Re: Kontiki]
#27017152 - 11/02/20 09:41 AM (3 years, 2 months ago) |
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Quote:
Kontiki said: Hey guys, I just made my first Agar a couple of days ago and it went quite well  After cooling a bit I took some tissue pieces from a B aspiring to get some nice clones, could anyone give me some help/how to think now when I’m about to make the first isolation. Which plate/ which part do I choose for the first transfer?

Cheers all!
You usually isolate as soon as possible. In your case now.
I'd isolate from 2 (nice rhizomorphic) and 5 (very uniform).
Were those tissues taken the same mushroom? And if so were the pieces close to each other?
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Log for my 2nd Grow (with pix and chronology) To shut up GIFs in Firefox: Go to about:config and set 'image.animation_mode' to 'none'. Then restart Firefox. For any other browser (especially Chrome): Uninstall browser. Switch to Firefox. See above.
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Kontiki
Stranger

Registered: 09/12/19
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Re: pour agar TEK [Re: funky123]
#27017190 - 11/02/20 10:00 AM (3 years, 2 months ago) |
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Aha I’ll get the SAB ready straight away. #1,2,3,4 is from the same mushroom, the inside of the lower part of the stem, with maybe a cm between each tissue sample. How come?
How is it, should I for example just take a tiiiiny piece of the outer part of the rhizo growth and put into a new petri dish? Or does it not matter which part I transfer?
What is the reason for your example to do 2 transfers with rhizo and 5 with uniform? 
Thanks for the help so far, just trying to understand this better )
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funky123
Pilzfreund


Registered: 08/27/17
Posts: 257
Loc: Austria
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Re: pour agar TEK [Re: Kontiki]
#27017401 - 11/02/20 12:27 PM (3 years, 2 months ago) |
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Quote:
Kontiki said: Aha I’ll get the SAB ready straight away. #1,2,3,4 is from the same mushroom, the inside of the lower part of the stem, with maybe a cm between each tissue sample. How come?
How is it, should I for example just take a tiiiiny piece of the outer part of the rhizo growth and put into a new petri dish? Or does it not matter which part I transfer?
What is the reason for your example to do 2 transfers with rhizo and 5 with uniform? 
Thanks for the help so far, just trying to understand this better )
Frankly speaking - I don't know for sure. I'm wondering myself about some of the things you are asking and didn't find good answers so far.
I mean technically speaking - I assume that if 1-4 are samples from the same mushroom the resulting mycelia should be genetically identical (as they are cloned from the same mushroom) - from which would follow that it doesn't matter where you isolate mycelium in 1 to 4.
I'd love to hear some educated opinion on that.
With spores it's a different story - there it makes sense to use a piece close to the perimeter and far from the central inoculation point as this will increase chances to isolate a genetically dominant mycelium.
> What is the reason for your example to do 2 transfers with rhizo and 5 with uniform? 
The reason is somewhere on spectrum from common sense to superstition. Good looks indicate health ...
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Log for my 2nd Grow (with pix and chronology) To shut up GIFs in Firefox: Go to about:config and set 'image.animation_mode' to 'none'. Then restart Firefox. For any other browser (especially Chrome): Uninstall browser. Switch to Firefox. See above.
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A.k.a
Stranger



Registered: 10/27/19
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Re: pour agar TEK [Re: funky123]
#27017472 - 11/02/20 01:00 PM (3 years, 2 months ago) |
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The reason for transferring the edge of the mycelium is because that’s the fastest moving newest part.
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LAGM2020     
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funky123
Pilzfreund


Registered: 08/27/17
Posts: 257
Loc: Austria
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Re: pour agar TEK [Re: A.k.a]
#27017515 - 11/02/20 01:17 PM (3 years, 2 months ago) |
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Quote:
A.k.a said: The reason for transferring the edge of the mycelium is because that’s the fastest moving newest part.
Sounds like an interesting idea but I'm not sure if I find that biologically plausible.
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Log for my 2nd Grow (with pix and chronology) To shut up GIFs in Firefox: Go to about:config and set 'image.animation_mode' to 'none'. Then restart Firefox. For any other browser (especially Chrome): Uninstall browser. Switch to Firefox. See above.
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