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Offlineta567445
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What is on my agar?
    #26678934 - 05/18/20 12:46 PM (3 years, 8 months ago)

Hi everyone one it is my first time going agar and I had a little bit of solution in a syringe left from a long time ago that I wanted to make viable again

I have 12 jars (mason jars no pour method)

Many/most of the jars look like this





Looks like contamination for me its been about 5 days since first inoculation

Where do I go from here, pardon my terminology, can I isolate the good looking growth/mycillium on to another agar wedge?

Any good teks on doing the transfer?

Thanks


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InvisibleModularMind
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Re: What is on my agar? [Re: ta567445]
    #26678941 - 05/18/20 12:48 PM (3 years, 8 months ago)

If you find good mycelium, transfer to a clean plate. How did you prepare your no pours?


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Offlineta567445
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Re: What is on my agar? [Re: ModularMind]
    #26678958 - 05/18/20 12:54 PM (3 years, 8 months ago)

Quote:

ModularMind said:
If you find good mycelium, transfer to a clean plate. How did you prepare your no pours?





Preparation
potato flake agar
honey
agar agar

PC @ 15 psi for 45 mins
125ml mason jars poly fill on the lids for breathing

SAB for inoculation


From the pics would you say taht I can just take the white parts and put it on a new plate?


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InvisibleRoger Clemency
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Re: What is on my agar? [Re: ta567445]
    #26678973 - 05/18/20 01:04 PM (3 years, 8 months ago)

Agar is meant for exactly that. Cutting out the good parts and transferring them away until you have just the good stuff. And it’s used for storing cultures and such.

But I don’t think the white you have there is cube mycelium. It could be some funny looking myc and the pictures making it look bad but I don’t know. Did you put spore solution all over the areas where the white is now?

Check out this link here- https://www.shroomery.org/forums/showflat.php/Number/18430998/fpart/1/vc/1


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InvisibleModularMind
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Re: What is on my agar? [Re: ta567445]
    #26679052 - 05/18/20 01:58 PM (3 years, 8 months ago)

Are they fluffy white or wet white spots?

I would leave the honey out and cut your PC time in half.
Especially for the shallow depth of media in no-pour form.


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Offlineta567445
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Re: What is on my agar? [Re: Roger Clemency]
    #26679066 - 05/18/20 02:05 PM (3 years, 8 months ago)

Quote:

Roger Clemency said:
Agar is meant for exactly that. Cutting out the good parts and transferring them away until you have just the good stuff. And it’s used for storing cultures and such.

But I don’t think the white you have there is cube mycelium. It could be some funny looking myc and the pictures making it look bad but I don’t know. Did you put spore solution all over the areas where the white is now?

Check out this link here- https://www.shroomery.org/forums/showflat.php/Number/18430998/fpart/1/vc/1





Thanks I read that thread but I am confused about wether my mycelium is good for extraction.... When I first inoculated I tried to make zig zag patter but the drops of the solution may have rolled around.


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Offlineta567445
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Re: What is on my agar? [Re: ModularMind]
    #26679072 - 05/18/20 02:07 PM (3 years, 8 months ago)

Quote:

ModularMind said:
Are they fluffy white or wet white spots?

I would leave the honey out and cut your PC time in half.
Especially for the shallow depth of media in no-pour form.




If i cut the honey out I can just use potato flakes and agar agar mix?

Its a fluffy white spot.


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InvisibleModularMind
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Re: What is on my agar? [Re: ta567445]
    #26679107 - 05/18/20 02:34 PM (3 years, 8 months ago)

I thought potato dextrose agar. My bad.
Most recipes call for dextrose.
Maybe boil your media once mixed and strain through cheese cloth or the like, to remove any solids, before pouring into jars for sterilization.

Do you have clean control jars from that batch?


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Offlineta567445
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Re: What is on my agar? [Re: ModularMind]
    #26679759 - 05/18/20 08:10 PM (3 years, 8 months ago)

Quote:

ModularMind said:
I thought potato dextrose agar. My bad.
Most recipes call for dextrose.
Maybe boil your media once mixed and strain through cheese cloth or the like, to remove any solids, before pouring into jars for sterilization.

Do you have clean control jars from that batch?





didnt have dextrose so used a substitute.

I didn't know I had to keep control jars.. I dont have any. What is the purpose of these?


Right now my only option is to some how clean the agar on new plates because I don't have any more spores left. Can you advise on where to start...

I am confused about wether to just do the extraction now or wait a couple of days for the myc to grow more in the jars (only been 5 days)


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InvisibleApples in Mono
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Re: What is on my agar? [Re: ta567445]
    #26679774 - 05/18/20 08:17 PM (3 years, 8 months ago)

Transfer a very small piece/pieces of the clean looking white mycelium as soon as you can


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Offlineta567445
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Re: What is on my agar? [Re: Apples in Mono]
    #26680965 - 05/19/20 11:57 AM (3 years, 8 months ago)

Quote:

Apples in Mono said:
Transfer a very small piece/pieces of the clean looking white mycelium as soon as you can





Thanks Apples


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Offlineta567445
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Re: What is on my agar?/progress [Re: ta567445]
    #26687602 - 05/22/20 08:34 AM (3 years, 8 months ago)



https://imgur.com/a/uXOrcWx

I took wedges out of my first cleanest looking batch and transferred to 7 new plates 6 of which look like the image attached i do see some rhizomorphic growth on this new one I opened for pics. (it has been 3 days since the new plates)

I do see green growth and black growth. Can someone advise me on where to go from here?

I am thinking of taking from the cleanest looking one and doing another 5 or 6 batch

I also noticed that my original plates look very clean now all the contaminated spots look like they have been taken over my mycelium. So should can I use those to put on new agars even though they looked contaminated at one point. The first mycelium plates have very fluffy dense growth vs any rhizomorphic features on this second one.

EDIT: the first one looks like the link below but when I scratched it looks like there is black spots under the mycelium you can see below.

https://imgur.com/a/PsB1NYU


Edited by ta567445 (05/22/20 09:24 AM)


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Offlineta567445
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Re: What is on my agar? [Re: Apples in Mono]
    #26687858 - 05/22/20 10:48 AM (3 years, 8 months ago)

Quote:

Apples in Mono said:
Transfer a very small piece/pieces of the clean looking white mycelium as soon as you can




I did the transfer see above.


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InvisibleRoger Clemency
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Re: What is on my agar? [Re: ta567445]
    #26687935 - 05/22/20 11:27 AM (3 years, 8 months ago)

So that first pic up top shows nice myc in the middle with satellite colonies of different molds around the plate. This means that when you’re doing your transfer to that plate you are letting other spores drop onto it. Either that or your filters on those jars is inadequate. You have to roll Polyfil very tight.

Are you doing these transfers in a sab? When you lift the lid make sure it’s a smooth motion and you don’t put your hand, arm, or anything but sterile scalpel blade over top of the plate. I lift lids as if they are hinged on the right side (lifting with right hand) and then move away an inch or so.

You can cut a piece of that good myc shown in the first pic. 12 o’clock right between the mold looks good but make it as far from them as possible and only a small sliver. I’d take another piece from about 5 o’clock too.

As far as the second picture, you don’t want to let the myc grow out that much before transferring since you really can’t see what’s underneath. When you know you have contaminant molds, or you know you’re going to transfer no matter what, always do it sooner than later. Get verifiably clean myc that you can see, grab it out and transfer. It can be 2-3 days sometimes after a transfer before I am doing another one.

Also, I don’t know much about molds except that there’s something called “spotted black mold” that is dangerous. Maybe deadly if breathed in. I don’t know for sure that’s what’s on the first pic but it’s a bunch of black spots so maybe look that up and be careful.


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Offlineta567445
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Re: What is on my agar? [Re: Roger Clemency]
    #26688000 - 05/22/20 11:49 AM (3 years, 8 months ago)

Quote:

Roger Clemency said:
So that first pic up top shows nice myc in the middle with satellite colonies of different molds around the plate. This means that when you’re doing your transfer to that plate you are letting other spores drop onto it. Either that or your filters on those jars is inadequate. You have to roll Polyfil very tight.

Are you doing these transfers in a sab? When you lift the lid make sure it’s a smooth motion and you don’t put your hand, arm, or anything but sterile scalpel blade over top of the plate. I lift lids as if they are hinged on the right side (lifting with right hand) and then move away an inch or so.

You can cut a piece of that good myc shown in the first pic. 12 o’clock right between the mold looks good but make it as far from them as possible and only a small sliver. I’d take another piece from about 5 o’clock too.

As far as the second picture, you don’t want to let the myc grow out that much before transferring since you really can’t see what’s underneath. When you know you have contaminant molds, or you know you’re going to transfer no matter what, always do it sooner than later. Get verifiably clean myc that you can see, grab it out and transfer. It can be 2-3 days sometimes after a transfer before I am doing another one.

Also, I don’t know much about molds except that there’s something called “spotted black mold” that is dangerous. Maybe deadly if breathed in. I don’t know for sure that’s what’s on the first pic but it’s a bunch of black spots so maybe look that up and be careful.





Thanks for the clear and concise answer. I am going to roll the polyfill super tight I wasn't rolling it tight before i thought it would completely stop air exchange... but also how tight is too tight?

I am using a SAB i'll be care full about the lids, I have been opening it fully for transfer . Ill do the ajar method.


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InvisibleRoger Clemency
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Re: What is on my agar? [Re: ta567445]
    #26688024 - 05/22/20 12:00 PM (3 years, 8 months ago)

I do take the lids fully off. My description isn’t exactly accurate, I lift the lid like it’s on a hinge and as it’s coming up I also move away from the plate. If you just do like a hinge you’d have your thumb and fingers hovering kinda over top of the lid.

Sorry if that’s just more confusing lol

I don’t think you can get poly too tight, especially for a Petri type that needs minimal GE


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Heaven starts right where hell ends


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