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eLeSDenes
Mycelium Expander




Registered: 05/25/11
Posts: 955
Last seen: 2 years, 7 days
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Invisible mycelium on agar
#18914527 - 10/01/13 03:40 AM (10 years, 4 months ago) |
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Hello everyone,
Sorry for making a new post but i did not find anything in the search engine. I have this problem for quite a long time. So whenever I do agar work, the mycelium looks very weak and almost invisibe. I think they are fine but i can only see the mycelium if i put the petri under a light. I have tried PDA, PDYA, MEA, MEYA, followed the recipes and tried to put just half or even quarter amount of nutrients. I always have to same result: almost transparent mycelium. I work in front of a HEPA so i got no problem with sterility. I looked some agar work of other members here and they got strong mycelium which is visible even without light. What am i doing wrong? And also if im making clones on agar i've never had any rhizo growth on my agar. I see some sectoring and tried to transfer a few times but i never get rhizo looking mycelium. Can anyone help me out? I would really appreciate that! Thank you:)
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SupaThaRipper
Genetics Hoarder



Registered: 09/02/13
Posts: 1,629
Loc: USA
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Re: Invisible mycelium on agar [Re: eLeSDenes]
#18914581 - 10/01/13 04:20 AM (10 years, 4 months ago) |
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Will someone answer this question already!? I am wondering as well
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RogerRabbit
Bans for Pleasure



Registered: 03/26/03
Posts: 42,214
Loc: Seattle
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Quote:
So whenever I do agar work, the mycelium looks very weak and almost invisibe.
Monokaryotic mycelium from spores will often be transparent. After a week or so you'll see it better as the strands thicken up after it becomes dikaryotic. RR
-------------------- Download Let's Grow Mushrooms semper in excretia sumus solim profundum variat "I've never had a failed experiment. I've only discovered 10,000 methods which do not work." Thomas Edison
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eLeSDenes
Mycelium Expander




Registered: 05/25/11
Posts: 955
Last seen: 2 years, 7 days
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Re: Invisible mycelium on agar [Re: RogerRabbit]
#18915420 - 10/01/13 10:16 AM (10 years, 4 months ago) |
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Thanks RR! I thought that monokaryotic mycelium mates just shortly after germination. I had almost fully colonized plates which looked transparent as well. I just remembered you saying that transfers from swipe should be done just after the first signs of growth. In that case i would just end up with a bunch of monokaryotic mycelium which still have to mate. Correct me if what im saying is wrong.
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Imperfect Iam
^means imperfect,not I'm perfect



Registered: 03/05/13
Posts: 7,237
Loc: center of the universe
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Re: Invisible mycelium on agar [Re: eLeSDenes]
#18966175 - 10/11/13 10:31 PM (10 years, 4 months ago) |
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I would love to know the answer to this also, as I get the same results from clones on agar, that I have been transferring as soon as it spread out enough so I could get a piece to transfer! I put some in grains as a matter of fact, two from same petri, which cont. with green cont. of some sort must have been in the plate, it was turkey tail, and it was the first one I did in Sab. Now I am afraid the other 4 are going to get the same shit, even thought I flame sterilized scalpel everytime. I know time will tell but would love to know sooner, as I asked a question about should I reclean box after moving on to another petri, but did not get a whole lot of info so I didn't!
-------------------- All you touch, and all you see, is all your life will ever be- Pink Floyd Life is what happens while you're busy making other plans- John Lennon
Edited by Imperfect Iam (10/13/13 02:36 AM)
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spiz
Stranger

Registered: 05/17/11
Posts: 118
Loc: Australia
Last seen: 2 years, 2 months
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Re: Invisible mycelium on agar [Re: RogerRabbit]
#24619970 - 09/11/17 06:55 AM (6 years, 5 months ago) |
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Quote:
RogerRabbit said:
Quote:
So whenever I do agar work, the mycelium looks very weak and almost invisibe.
Monokaryotic mycelium from spores will often be transparent. After a week or so you'll see it better as the strands thicken up after it becomes dikaryotic. RR
So at which point is it best to transfer? Its my first time working with agar and on one of my plates i have spores that have germinated and can see some sectoring. The growth is transparent and can only be seen when tilted to the light at the right angle.
In your videos they're very white before you transferred. Is that the point i get to before transferring or was that just done so it was clear for the video
Thank you
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Pipefitter537
You're Not Yelping



Registered: 11/25/16
Posts: 715
Loc: SodoSopa
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Re: Invisible mycelium on agar [Re: spiz]
#24619979 - 09/11/17 07:03 AM (6 years, 5 months ago) |
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If you can see that's it's clean growth, than transfer. If I'm starting from spores, I transfer early, or as soon as I can tell it's myc. Take a small piece and you should be be. Transfer further down the line will be cleaner, so you can let them grow further. I just don't let them grow out enough to touch the sides of the plate. Good luck.
-------------------- "You should never underestimate the predictability of stupidity"-Bullet Tooth Tony
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spiz
Stranger

Registered: 05/17/11
Posts: 118
Loc: Australia
Last seen: 2 years, 2 months
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Thanks for reply.
Even if I'm using a SAB? I may be able to see enough to transfer a chunk but there's absolutely no way I could see enough to transfer the different sectors.
Is transferring at such an early stage only really viable if you're using a flow hood?
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Pipefitter537
You're Not Yelping



Registered: 11/25/16
Posts: 715
Loc: SodoSopa
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Re: Invisible mycelium on agar [Re: spiz]
#24620095 - 09/11/17 08:25 AM (6 years, 5 months ago) |
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Nah. Don't worry about sectors yet. Sometimes they show right from germination but not always. Focus on clean, then sectors if you want to go that route.
I hold my plates or dishes to a light outside sab, make a mark with a sharpie where I want to take a piece from, then bring into sab and transfer said piece. Easier than looking at an open plate for too long to decide where to transfer from
-------------------- "You should never underestimate the predictability of stupidity"-Bullet Tooth Tony
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spiz
Stranger

Registered: 05/17/11
Posts: 118
Loc: Australia
Last seen: 2 years, 2 months
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So what's the point of transferring then? If there's no contamination (from an absolute newbies point of view) and you're not trying to get each sector yet then why bother transferring?
Not saying you're wrong or anything just trying to learn the motives behind the method.
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bodhisatta 
Smurf real estate agent


Registered: 04/30/13
Posts: 61,890
Loc: Milky way
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Re: Invisible mycelium on agar [Re: spiz]
#24620157 - 09/11/17 08:53 AM (6 years, 5 months ago) |
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Quote:
spiz said: So what's the point of transferring then? If there's no contamination (from an absolute newbies point of view) and you're not trying to get each sector yet then why bother transferring?
Not saying you're wrong or anything just trying to learn the motives behind the method.
From spores make 1-2 transfers or more as needed to get clean growth then roll with it.
Clone and keep clones around to do awesome grows
Do isolate work in spare time, isolate work is blind, you have to test each one for performance since you cant tell anything based on how it looks on agar. Most suck so make at least ten to test at a time
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spiz
Stranger

Registered: 05/17/11
Posts: 118
Loc: Australia
Last seen: 2 years, 2 months
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Re: Invisible mycelium on agar [Re: bodhisatta]
#24620181 - 09/11/17 09:01 AM (6 years, 5 months ago) |
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Ahhh right. I thought you wanted an isolate before trying to colonize grain because it colonized faster or something.
Is that correct? Is getting isolates more for really advanced growers?
Thank you both for your help
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Pipefitter537
You're Not Yelping



Registered: 11/25/16
Posts: 715
Loc: SodoSopa
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Re: Invisible mycelium on agar [Re: bodhisatta]
#24620189 - 09/11/17 09:03 AM (6 years, 5 months ago) |
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Beat me too it. Once you start agar you will have a shitload of plates with various projects going, cloning, germination, etc. I Like bod said, isolate search is a good side project while other shits doing its thing
As long as you have clean growth, it will colonize quick. And depending on how much of the plate you noc the jars with, the colonizing time will vary, the more used the faster and the less the slower.
-------------------- "You should never underestimate the predictability of stupidity"-Bullet Tooth Tony
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spiz
Stranger

Registered: 05/17/11
Posts: 118
Loc: Australia
Last seen: 2 years, 2 months
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Thank you guys so much!!
I think you just saved me a lot of unnecessary time wasted
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bodhisatta 
Smurf real estate agent


Registered: 04/30/13
Posts: 61,890
Loc: Milky way
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Re: Invisible mycelium on agar [Re: spiz]
#24620296 - 09/11/17 09:50 AM (6 years, 5 months ago) |
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I almost never grow out isolates. Almost everything i do is MS or a clone
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spiz
Stranger

Registered: 05/17/11
Posts: 118
Loc: Australia
Last seen: 2 years, 2 months
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Re: Invisible mycelium on agar [Re: bodhisatta]
#24620328 - 09/11/17 10:09 AM (6 years, 5 months ago) |
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I could kiss you right now hahaha
After you guys set me in the right direction I was able to start searching for the right things.
I now realise the "sectoring" I see is just multiple spores germinating and that finding an isolated takes a lot of effort. I literally labelled the plate I was referring to as 'visible' because I scratched a shit ton of spores onto it to make sure something would grow
Again thank you for clearing things up for me you guys are legends
Edited by spiz (09/11/17 10:11 AM)
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Pipefitter537
You're Not Yelping



Registered: 11/25/16
Posts: 715
Loc: SodoSopa
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Re: Invisible mycelium on agar [Re: spiz]
#24620556 - 09/11/17 11:45 AM (6 years, 5 months ago) |
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Check bods links for his agar shit. Great read.
Here Is another great thread https://www.shroomery.org/forums/showflat.php/Number/24330957
Here is a great isolate read if you ever go that route https://www.shroomery.org/forums/showflat.php/Number/18430998
-------------------- "You should never underestimate the predictability of stupidity"-Bullet Tooth Tony
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