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Re: AGAR ENVY! (Anything and All things agar!) [Re: AspectOfTheCreator] * 3
    #28885956 -

I just swirl and pour hot as balls.

If you use good ingredients that kind of shit doesn't happen. Go for light pilsen malt like lovibond 1 or 2. Mix dry ingredients with water in your bottle of choice and pc. Swirl when you can handle it safely and pour.



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

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Re: AGAR ENVY! (Anything and All things agar!) [Re: Nichrome]
    #28886046 -

what is that?


--------------------




" Antes de que cambien los reinos, es necesario que cambien los individuos"

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Re: AGAR ENVY! (Anything and All things agar!) [Re: DRAKENCL] * 3
    #28886069 -

Bacteria.


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

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Re: AGAR ENVY! (Anything and All things agar!) [Re: Nichrome] * 3
    #28886990 -

These TTBVI are flying! I read that they are fast, but I wasn't expecting this haha. Two days (but less than 48 hours) since last pictures:





Looking super clean thanks to SupaThaRipper! Well done, man :mushroom2:



A T2 Yeti soon to move into a jar:



A T1 Yeti that's more rhizomorphic than the T2; might also be sent to Jarlandia for the fun of it:




And then this interesting growth; I've never had a plate do this:





:shrug: Might be aliens.

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Re: AGAR ENVY! (Anything and All things agar!) [Re: ElVatoFirme]
    #28887055 -

I need some help with my agar. I have recently started doing agar work in a SAB and I've gotten some nice cultures to grow out and now have tubs about to fruit from them, so I've had some success. But I've been having a very high rate of bacterial contam coming in around the outsides of my plates and anywhere the water inside contacts the agar.

I was thinking it was because of condensation so I tried to reduce that by pouring at lower temps. And that helped reduce the condensation by a lot but I still get some and I still get contams from it. It seems like its impossible to completely eliminate condensation, unless you have a flow hood you can dry them in front of before sealing.


I've been using the parafilm grafting tape to seal my plates, instead of the sterile laboratory grade tape, because its a lot cheaper and I've seen several anecdotal reports of it working fine. Is this my problem? Maybe the bacteria is being introduced to the condensation by this tape?

Can anybody offer any tips to help eliminate condensation, and/or to stop it from becoming bacterial? TIA

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Re: AGAR ENVY! (Anything and All things agar!) [Re: zorak]
    #28887071 -

Condensation alone shouldn't be causing contamination.

I wonder if somehow when you're lifting or replacing the lid, you're touching water with your hands that later falls onto the plate. Might be worth being really careful to lift the lid without touching the bottom rim of the lid.

Parafilm isn't really sterile, especially after we stretch it with our fingers, so I doubt changing the wrapper you're using would matter.

Are you seeing contamination on blank plates? Are you wrapping blank plates, or storing them in a tote unwrapped?


--------------------
LAGM 2026 grow log | My library/trade list

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Re: AGAR ENVY! (Anything and All things agar!) [Re: Phaethon]
    #28887085 -

Yes, I made a bunch of blank plates and wrapped and stored them, almost all contaminated. Even with a lot of plates that had good myc growth, it only grow out until it reached a ring of contam around the outside.

I'm sure I have been touching the bottom rim of the lid, I have not been trying to avoid that. But I wear gloves sanitized with iso so I thought I would be safe to touch it there without causing any problems. Is that not true?

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Re: AGAR ENVY! (Anything and All things agar!) [Re: zorak]
    #28887092 -

zorak said:
Yes, I made a bunch of blank plates and wrapped and stored them, almost all contaminated. Even with a lot of plates that had good myc growth, it only grow out until it reached a ring of contam around the outside.

I'm sure I have been touching the bottom rim of the lid, I have not been trying to avoid that. But I wear gloves sanitized with iso so I thought I would be safe to touch it there without causing any problems. Is that not true?


Hey Zorak

Plates contaminating before using them needs to be addressed first. Best not to do any transfers until you can make clean plates

Touching the bottom rim is an invitation for contamination. So, yeah, best to avoid that
Edit: using the clamshell technique is a really good skill

Nice job seeking the causes

Oh, still looking forward to pics of that tub!


--------------------
Hitchhiker’s Guide to the Shroomery “Clean spawn is easily the most critical component of the entire cultivation process.”
Drink the Water of Life and become Paul Muad’Dib.
Remember to surrender.

Edited by AzureCultivator (07/25/24 06:38 PM)

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Re: AGAR ENVY! (Anything and All things agar!) [Re: zorak]
    #28887093 -

zorak said:
Yes, I made a bunch of blank plates and wrapped and stored them, almost all contaminated. Even with a lot of plates that had good myc growth, it only grow out until it reached a ring of contam around the outside.

I'm sure I have been touching the bottom rim of the lid, I have not been trying to avoid that. But I wear gloves sanitized with iso so I thought I would be safe to touch it there without causing any problems. Is that not true?


Is it possible that your agar has gotten contaminated somehow and you need to mix up some other?

If you are wiping down your whole sab
shutting down all fans
using clean gloves
changed bottles you mix it in

it almost has to be something else?

I lived down in summit for almost a year and could not keep a clean jar. I had to switch to tyndallizing to get 98% and i never figured out if it was the moldy place i worked in or the grain or a dam hateful poltergeist


--------------------
Yours in the Natural State Land of Enchantment!

"Sometimes the only way for a man to find true happiness is to take drugs in a group" - Cochise

No makin funna my pomes!

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Re: AGAR ENVY! (Anything and All things agar!) [Re: zorak] * 1
    #28887192 -

zorak said:
Yes, I made a bunch of blank plates and wrapped and stored them, almost all contaminated. Even with a lot of plates that had good myc growth, it only grow out until it reached a ring of contam around the outside.

I'm sure I have been touching the bottom rim of the lid, I have not been trying to avoid that. But I wear gloves sanitized with iso so I thought I would be safe to touch it there without causing any problems. Is that not true?


Isopropyl doesn't sterilize. If you wipe your gloved hands with iso and touch agar, you will almost certainly see bacterial and maybe moldy growth where you touched. Still, to be honest, I'm not super meticulous about how I pick up lids, or at least I don't think about it much anymore, and I don't get bacterial contamination around plate edges.

In addition to Azure's and curenado's responses, I'm a little suspicious of your dishes and maybe your agar prep.

Are you using disposable plastic dishes (which are sterile until you open them) or polypropylene or glass ones you have to sterilize yourself? If not pre-sterilized, how are you sterilizing? Are you wrapping inside the SAB?

How long are you sterilizing your media? I think 20 minutes is too short, though lots of people get away with it; I always do 45 minutes for 400 ml bottles of agar or LC broth. More is more, as Stipe likes to point out.


--------------------
LAGM 2026 grow log | My library/trade list

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Re: AGAR ENVY! (Anything and All things agar!) [Re: curenado]
    #28887215 -

I make a new batch for each pour. I sterilize it in an instant pot.

One thing that concerns me is the contaminated air that comes in as it cools inside the instant pot.... The whole point of turning the lid upside down is so air can escape as it heats up and enter as it cools, but what is stopping that cool air from contaminating the batch?


And idk how I could totally avoid touching the rim of the lid. I pour a stack of plates by starting on the bottom and going up. But when I lift the bottom lid and every plate above it, I need to have a pretty firm grip to prevent the top ones from falling off. So typically I'm touching a lot of the upper plates when I do this, and the bottom rim of the bottom plates lid. I guess this must be the problem based on what you're saying, but how do you pick up a whole stack without doing that?

Btw azure I just took some pics, will share probably later when I have more progress pics and results to post with it. Looks like the tub I posted in the other thread may have stalled a bit from the heavy misting, but the other 2 are progressing nicely and I'm sure the 1st will catch up

Edited by zorak (07/25/24 08:03 PM)

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Re: AGAR ENVY! (Anything and All things agar!) [Re: zorak] * 1
    #28887247 -

WEZA

MIB Red Spore

BISPORUS


Would like any opinions on how these look, i've only been doing agar for 5-6 months and this is my proper try at growing Pan. Species. they look beautiful to me. I'm enjoying agar work more than I do actually growing shrooms.


--------------------
Yummy!


LAGM 2025
https://www.shroomery.org/forums/showflat.php/Number/29174693#29174693

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Re: AGAR ENVY! (Anything and All things agar!) [Re: syllable]
    #28887300 -

The Thing





--------------------
I call them Huskies cause they Mush!


The Official Wild Cubensis Thread
Instant Pot Agar
Gnocchi Water Agar
Let's Talk Cowtek

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Re: AGAR ENVY! (Anything and All things agar!) [Re: zorak]
    #28887335 -

zorak said:
And idk how I could totally avoid touching the rim of the lid. I pour a stack of plates by starting on the bottom and going up. But when I lift the bottom lid and every plate above it, I need to have a pretty firm grip to prevent the top ones from falling off. So typically I'm touching a lot of the upper plates when I do this, and the bottom rim of the bottom plates lid. I guess this must be the problem based on what you're saying, but how do you pick up a whole stack without doing that?



Ah, I keep the plastic wrap on the stack. I only touch the top plate of each set. Start at the bottom, tiny lift and tilt, pour, replace and on to the next. Adjust the plastic wrap as I work my way up. I do about 8 at a time. More gets tricky

My way is weak compared to this: Pouring agar — YouTube


--------------------
Hitchhiker’s Guide to the Shroomery “Clean spawn is easily the most critical component of the entire cultivation process.”
Drink the Water of Life and become Paul Muad’Dib.
Remember to surrender.

Edited by AzureCultivator (07/25/24 10:10 PM)

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Re: AGAR ENVY! (Anything and All things agar!) [Re: AzureCultivator]
    #28887360 -

If I had to fiddle with plastic wrap while pouring agar I would never pour agar :rofl:


--------------------
LAGM 2026 grow log | My library/trade list

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Re: AGAR ENVY! (Anything and All things agar!) [Re: zorak]
    #28887374 -

zorak said:
I make a new batch for each pour. I sterilize it in an instant pot.

One thing that concerns me is the contaminated air that comes in as it cools inside the instant pot.... The whole point of turning the lid upside down is so air can escape as it heats up and enter as it cools, but what is stopping that cool air from contaminating the batch?


And idk how I could totally avoid touching the rim of the lid. I pour a stack of plates by starting on the bottom and going up. But when I lift the bottom lid and every plate above it, I need to have a pretty firm grip to prevent the top ones from falling off. So typically I'm touching a lot of the upper plates when I do this, and the bottom rim of the bottom plates lid. I guess this must be the problem based on what you're saying, but how do you pick up a whole stack without doing that?

Btw azure I just took some pics, will share probably later when I have more progress pics and results to post with it. Looks like the tub I posted in the other thread may have stalled a bit from the heavy misting, but the other 2 are progressing nicely and I'm sure the 1st will catch up


I'm sure my fingers are often touching the bottom of the rim of the lid when I lift a stack when pouring, too. I'm a little sorry for even bringing it up.

I doubt your contamination is due to air getting sucked in if you're leaving the lid a little loose. We all leave our lids loose to cool, and if you tighten too early, you have more danger of a fast "burp" when opening the jar in your SAB.

Could be you're touching the rim of the jar when removing the flat lid before pouring...but I'd expect to see contamination elsewhere than the edges of the plates. Still if you have one piece polypropylene lids, it's worth trying those instead. Or a media bottle (the 250 ml ones fit in a 6 qt Instant Pot). Those have these magical rings that prevent any dripping or running down the side when pouring.

Are you moving the stack so it's not hovering right above the open dish as you're pouring?

And how about the pressure cook time (I asked earlier but might have gotten buried)? And are you venting steam for 10 minutes before putting it in seal mode?

Sorry, really no smoking gun here so I'm just asking lots of questions in case one of the answers turns up the culprit.


--------------------
LAGM 2026 grow log | My library/trade list

Edited by Phaethon (07/25/24 10:41 PM)

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Re: AGAR ENVY! (Anything and All things agar!) [Re: Phaethon]
    #28887395 -

I move the stack as little as possible so it is still partially hovering over. But I think the fact that it shows up as a ring around the outside means the agar isn't contaminated before pouring. I'm pretty sure the condensation is the vector and the bacteria is making it's way in from outside the dish. 
I do have a tendency to flip them around when looking at them which I guess might be the cause? cause when its flipped upside down the condensation runs into the bottom of the lid and then and when it's flipped upright again, it can run down the sides of the lid and into the little space under the bottom lip. The grafting tape generally seals it pretty well so it doesn't make it out from there but I guess that could be where its picking up bacteria. Maybe I just need to be careful about not flipping it more than once?

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Re: AGAR ENVY! (Anything and All things agar!) [Re: zorak]
    #28887408 -

I keep plates in ziplocks now. Makes a big difference. Look but don’t touch


--------------------
Hitchhiker’s Guide to the Shroomery “Clean spawn is easily the most critical component of the entire cultivation process.”
Drink the Water of Life and become Paul Muad’Dib.
Remember to surrender.

Extras: Filter Print Post Top
Re: AGAR ENVY! (Anything and All things agar!) [Re: Phaethon]
    #28887413 -

Phaethon said:
zorak said:
Yes, I made a bunch of blank plates and wrapped and stored them, almost all contaminated. Even with a lot of plates that had good myc growth, it only grow out until it reached a ring of contam around the outside.

I'm sure I have been touching the bottom rim of the lid, I have not been trying to avoid that. But I wear gloves sanitized with iso so I thought I would be safe to touch it there without causing any problems. Is that not true?


Isopropyl doesn't sterilize. If you wipe your gloved hands with iso and touch agar, you will almost certainly see bacterial and maybe moldy growth where you touched. Still, to be honest, I'm not super meticulous about how I pick up lids, or at least I don't think about it much anymore, and I don't get bacterial contamination around plate edges.

In addition to Azure's and curenado's responses, I'm a little suspicious of your dishes and maybe your agar prep.

Are you using disposable plastic dishes (which are sterile until you open them) or polypropylene or glass ones you have to sterilize yourself? If not pre-sterilized, how are you sterilizing? Are you wrapping inside the SAB?

How long are you sterilizing your media? I think 20 minutes is too short, though lots of people get away with it; I always do 45 minutes for 400 ml bottles of agar or LC broth. More is more, as Stipe likes to point out.


Just to say I don't run 20 minutes either. I run 30 minutes on 250ml and funny enough, the same on pints as you, just trying to avoid bummers.


--------------------
Yours in the Natural State Land of Enchantment!

"Sometimes the only way for a man to find true happiness is to take drugs in a group" - Cochise

No makin funna my pomes!

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Re: AGAR ENVY! (Anything and All things agar!) [Re: zorak]
    #28887437 -

zorak said:
I need some help with my agar. I have recently started doing agar work in a SAB and I've gotten some nice cultures to grow out and now have tubs about to fruit from them, so I've had some success. But I've been having a very high rate of bacterial contam coming in around the outsides of my plates and anywhere the water inside contacts the agar.

I was thinking it was because of condensation so I tried to reduce that by pouring at lower temps. And that helped reduce the condensation by a lot but I still get some and I still get contams from it. It seems like its impossible to completely eliminate condensation, unless you have a flow hood you can dry them in front of before sealing.


I've been using the parafilm grafting tape to seal my plates, instead of the sterile laboratory grade tape, because its a lot cheaper and I've seen several anecdotal reports of it working fine. Is this my problem? Maybe the bacteria is being introduced to the condensation by this tape?

Can anybody offer any tips to help eliminate condensation, and/or to stop it from becoming bacterial? TIA


Do you have a rack in your SAB that you work on or are you pouring on the floor of it?


--------------------
𝕴𝖋 𝕴 𝖍𝖆𝖛𝖊 𝖘𝖊𝖊𝖓 𝖋𝖚𝖗𝖙𝖍𝖊𝖗 𝖎𝖙 𝖎𝖘 𝖇𝖞 𝖘𝖙𝖆𝖓𝖉𝖎𝖓𝖌 𝖚𝖕𝖔𝖓 𝖙𝖍𝖊 𝖘𝖍𝖔𝖚𝖑𝖉𝖊𝖗𝖘 𝖔𝖋 𝕲𝖎𝖆𝖓𝖙𝖘
:drugnerd: TekDeck        Autist Tally :laugh:

"Mother Mycelium, I wandered in the darkness of unmarked plates and forgotten codes. Enlighten me with clarity and discipline, that every jar and dish shall bear its true name."

"Luminous Logos, I faltered in my shorthand faith. Grant me the diligence to record in full, and the foresight to recall every meaning with certainty."

-Father LewDoja

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