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Re: AGAR ENVY! (Anything and All things agar!) [Re: Tri]
    #28887506 -

I believe I pc'd for 30 minutes generally, plus a 10 minute vent. I've reduced it to 25 mins once when I accidentally did a 20 minute vent. I didn't want to go too long and caramelize it. I dont use a rack in my SAB.

Now that I think about it I've even had some of my unused no-pour cups go bacterial. But I had some from the same batch that were ok. Makes me think it's from contaminants in the air that come in while the instant pot is cooling. I actually put a bleach-soaked towel over the vent hole ast it was cooling last time, hoping to kill anything coming in. But the heat of the lid probably made it evaporate off really quickly

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Re: AGAR ENVY! (Anything and All things agar!) [Re: zorak] * 1
    #28887515 -

I really doubt it's dirty air coming into your Instant Pot and through the threads of your lid that's causing this. For one thing, during most of thr cooling time, temperatures are high enough to instantly kill anything other than endospores, which are not usually airborne. And I don't think many people are filtering their pressure cooker vents.

If you have unused no pours going bacterial, then I'd guess you are under-sterilizing. And since most Instant Pot models don't get above 12 psi, I wonder if 30 minutes is sufficient.


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Re: AGAR ENVY! (Anything and All things agar!) [Re: zorak]
    #28887566 -

That makes sense. I guess I will have to try cooking it longer. Although I believe I was already adding time to the instant pot tek I found which seemed to work fine for other people. If I go for more than 45 minutes then it caramelizes and becomes unusable to the mycelium, right? I guess I'll try 40

Edit - I make mea not pda, if that matters

Edited by zorak (07/26/24 04:59 AM)

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Re: AGAR ENVY! (Anything and All things agar!) [Re: zorak] * 1
    #28887585 -

Nah. 45min is fine for agar. I always vent for 20, cycle for 45 in my instant pot. The sediment that may form won't hurt anything, it's just ugly. I sometimes let em sit on "keep warm" for a while to let junk settle to bottom, but it's purely aesthetic. Other folks stir it all back together before pouring.


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This hobby has taught me many things.
Most of those things are patience.

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Re: AGAR ENVY! (Anything and All things agar!) [Re: Scemilope] * 1
    #28888509 -

TY I wil definitely try that! I just remembered, I think I gave my last batch a little less time in the PC because I was trying to get perfectly clear and pretty looking plates. Then when most of them contaminated I got discouraged and almost gave up on Agar. But I will definitely try it again with a longer cook time and forget about the sediment.

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Re: AGAR ENVY! (Anything and All things agar!) [Re: zorak] * 2
    #28888510 -

zorak said:
TY I wil definitely try that! I just remembered, I think I gave my last batch a little less time in the PC because I was trying to get perfectly clear and pretty looking plates. Then when most of them contaminated I got discouraged and almost gave up on Agar. But I will definitely try it again with a longer cook time and forget about the sediment.



Sediment can be curbed with purified water. Try distilled.


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Astra said:

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Re: AGAR ENVY! (Anything and All things agar!) [Re: TheAtlantean]
    #28888582 -

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

 
Do you wrap them first? And if not, how fast do they dry?

My last batch had very little condensation, but it still took about 3-4 weeks for that little bit to dry out with them sitting out on a shelf, wrapped in grafting tape. I was expecting them to dry faster than that.

TheAtlantean said:
zorak said:
TY I wil definitely try that! I just remembered, I think I gave my last batch a little less time in the PC because I was trying to get perfectly clear and pretty looking plates. Then when most of them contaminated I got discouraged and almost gave up on Agar. But I will definitely try it again with a longer cook time and forget about the sediment.



Sediment can be curbed with purified water. Try distilled.


Ok I will try distilled next time too, I've been using RO.

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Re: AGAR ENVY! (Anything and All things agar!) [Re: zorak]
    #28888668 -

Don't worry about caramalisation, I've cooked my agar for an hour in the IP and I've run it through 2 x 45 min cycles in a row because I forgot I did the first cycle without any issues.

I also wouldn't worry too much about some condensation, unless it's pooling on the surface of the agar, it doesn't effect performance and you can clear condensation with a flat bottom mug on top of the plate for a couple mins to view the plate.

I let my plates cool and use any I need immediately, and then put the bag the empty plates came in back over the stack and tape the bottom up :shrug:

I'll wrap plates when I next open the bag to use plates or if the stack of plates sits for more than a week in the bag unwrapped.

I highly recommend using clingfilm to wrap plates, it's cheap and easy to work with, the only problem is cutting the roll which you can do with a pipe cutter or really sharp knife pretty easily.

I do 2 x wraps of Glad brand clingfilm around a plate, Glad seems to stick to plates better and feels better to work with IMO than cheap brands.

Are you mixing your agar ingredients with cold water to dissolve dry malt extract and avoid the agar clumping, and then melting the agar, and mixing again before putting it through the IP cycle?

I put my ingredients directly into my lab bottle, put the bottle onto a magnetic stirrer to dissolve dry malt extract and then put my lab bottle in a hot water bath to melt ingredients, back onto magnetic stirrer again for a min, 45 min IP cycle with a 5 min vent, onto magnetic stirrer again once it's done and then pour once it's cool enough to handle.

I used to shake instead of using the magnetic stirrer and it came out the same, but needed to wait for the bubbles to disappear before pouring.



This plate is tap water, 1% DLME and 2% agar and our tap water is considered "hard", but I can clearly see through my agar and haven't noticed any sediment.

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Re: AGAR ENVY! (Anything and All things agar!) [Re: Dkeppel]
    #28889360 -

Thanks again to everyone. Will keep trying. I'm about to order more petri dishes and I'm wondering if I could've gotten bad ones last time... Every listing on amazon has a few bad reviews claiming they were contaminated, but I'm assuming most of those people are just blaming the dishes for their bad technique. 
Is there a certain brand that people can recommend? Or brands that should be avoided?


Btw the contamination I'm talking about is very difficult to see, it just looks like the surface of the agar changes to look less glossy and smooth, but you have to hold it in just the right light to be able to see it. For a long time I was unsure if it was actually contam, or just changes in the surface of the agar due to moisture or some other reason. But my cultures stop growing when they hit it so it must be contam. Just wanted to mention that in case it indicates anything.

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Re: AGAR ENVY! (Anything and All things agar!) [Re: zorak]
    #28889380 -

Sorry if I missed it, but what plates are you using? Glass/plastic/sauce containers? There is a difference in the workflow so that matters. Other than that, I would blame your sterile technique if you're getting bacterial spots on your plates.

It's difficult to be certain without photos, but the way you're describing it does sound like bacteria colonizing some of the plate before the mycelium reaches it. Maybe share your complete transfer procedure step by step?

Placing your finger on the inside of the plate lid will almost certainly lead to contamination, so get rid of that first. Sanitization does not equal sterilization. Grab your plates on the outside edge of the lid and just hold them between your fingers. Make sure not to hover anything other than your flame sterilized scalpel over the plate.

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Re: AGAR ENVY! (Anything and All things agar!) [Re: zorak]
    #28889404 -

It is possible I guess, but I've used hundreds of plates from Temu and Aliexpress without any issue and that's probably where Amazon plates come from originally?

I think you mostly just need to run at least a 45min cycle and make sure to include at a minimum a 5 min vent by the sound of it.

It sounds like bacteria, probably those tough little endospore bastards surviving due to improper sterilisation.

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Re: AGAR ENVY! (Anything and All things agar!) [Re: Dkeppel]
    #28889412 -

I use plastic pre-sterilized petri dishes mostly. Also have made no-pour cups using small pc-able containers with screw-on lids

It doesn't show up in spots, it shows up as a ring around the outer diameter of the plate, and/or streaks where water has run across the surface.

Edited by zorak (07/27/24 09:17 PM)

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Re: AGAR ENVY! (Anything and All things agar!) [Re: zorak] * 1
    #28889779 -









Good afternoon everyone. These are my yeast agar plates: 1 gram of brewer's yeast, 20 grams of dextrose, 20 grams of agar, 20 grams of brown sugar. I notice yeast growth on some of these plates; today I transferred some portions to salvage this beautiful TAT.


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" 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]
    #28889962 -

:congrats:
Beautiful rhizos. I have never made a glass window agar but dang it's pretty!


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Yours in the Natural State Land of Enchantment!

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Re: AGAR ENVY! (Anything and All things agar!) [Re: curenado]
    #28890134 -

Shoutout to the creators and contributors of this thread…it’s been super helpful!

After a lot of trial and error…it’s nice to see some progress. Just have a couple of questions on the progress.

Aff Natalensis T2

Image 1: Tomentose growth or mold on the left and rhizomorph on right?



Image 2: Would this be considered “organised or clean” enough to send to grain?


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Re: AGAR ENVY! (Anything and All things agar!) [Re: salubrious]
    #28890148 -

salubrious said:
Shoutout to the creators and contributors of this thread…it’s been super helpful!

After a lot of trial and error…it’s nice to see some progress. Just have a couple of questions on the progress.

Aff Natalensis T2

Image 1: Tomentose growth or mold on the left and rhizomorph on right?

Image 2: Would this be considered “organised or clean” enough to send to grain?



Pic 1 - tomatose on left and rhizo on right.  Take transfers from each side and finish getting more symmetrical growth patterns before fruiting them seperately as they'll likely have different phenotypes expressed in the fruits.
Pic 2 - looks clean, but not ready for grain yet IMO.  Another transfer or two should do it though.

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Re: AGAR ENVY! (Anything and All things agar!) [Re: PBJ710]
    #28890207 -

I purchased some premade agar plates- what’s the best way to store them before adding some genetics and after- once the culture has grown?

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Re: AGAR ENVY! (Anything and All things agar!) [Re: Fighting_CRPS]
    #28890214 -

I just keep mine in a ziplock. No fridge or anything


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Re: AGAR ENVY! (Anything and All things agar!) [Re: AspectOfTheCreator]
    #28890217 -

Sweet! Thank you!

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Re: AGAR ENVY! (Anything and All things agar!) [Re: Fighting_CRPS]
    #28890957 -

should be good for a couple months at room temp ime, even longer in the fridge


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Mushrooms, Mycology and Psychedelics > Mushroom Cultivation
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