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 SirPsycho
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 Re: No-pour agar sucks!  [Re: SirPsycho] 2
#28100938 - 12/16/22 06:06 PM (1 year, 3 months ago) |
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As for OP, you're making at least two wrong assumptions.
First. Mycelium does not grow into the agar, only across the surface. I'm not gonna get into a debate about it again. This is literally the whole point of agar.
Sedond. As has been said, nutrients do not settle. They are dissolved into solution and evenly distributed.
On another note, my no pours have always contaminated when they should. For the most part I could always tell right at inoculation which plates would contam cause I fucked up.
Edit: I would look at your grain prep or transfer technique for A2G
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 Mycolorado
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Re: No-pour agar sucks! [Re: SirPsycho]
#28100944 - 12/16/22 06:09 PM (1 year, 3 months ago) |
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Just reread the op. In that case, the only thing I would think is that small aliquots of medium are being pcβd too long and denaturing/changing the nutrients via the Maillard reaction but that seems a stretch. I donβt believe nutrient settling and stratification is an issue as they are in solution unless itβs somehow related to the volume/pc time.
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SirPsycho
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I regularly pressure cook my no pours for 45 mins without any issues like caramelization or anything like that.
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Mycolorado
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Re: No-pour agar sucks! [Re: SirPsycho]
#28100950 - 12/16/22 06:16 PM (1 year, 3 months ago) |
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Hence why I said that would be a stretch.
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SirPsycho
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Yep, and I'm pretty sure I've seen p9 say sugars can't caramelize in the pc. Forget exactly why
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JHOVA
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Re: No-pour agar sucks! [Re: SirPsycho]
#28101014 - 12/16/22 07:15 PM (1 year, 3 months ago) |
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So if your agar on both fronts goes bad how will you use agar? Going back to spore syringe and pf tek? Whats your end goal???
Do you want help with no pour or are you just going to do pour agar? Trying to understand best i can
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SirPsycho
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Re: No-pour agar sucks! [Re: JHOVA]
#28101017 - 12/16/22 07:18 PM (1 year, 3 months ago) |
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Quote:
JHOVA said:
So if your agar on both fronts goes bad how will you use agar? Going back to spore syringe and pf tek? Whats your end goal???
Do you want help with no pour or are you just going to do pour agar? Trying to understand best i can
Quote:
SirPsycho said:
The issue here, at least as OP is putting it and with me just skimming the thread, he's ending up with contaminated grain spawn and can't identify the source. His running theory at this point is that there is contam on his no pour agar but it isn't germinating on the plates, so he can't see it on the plates but it then kicks off when putting that agar into grain.
So he's purposefully trying to contam his plates to prove that there's an issue with nasties not germinating on no pour.
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LadysKnight
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Re: No-pour agar sucks! [Re: SirPsycho]
#28101030 - 12/16/22 07:32 PM (1 year, 3 months ago) |
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JHOVA
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Re: No-pour agar sucks! [Re: SirPsycho]
#28101032 - 12/16/22 07:34 PM (1 year, 3 months ago) |
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Thank you for linking that. I get it now!
This is similar to Josex thread where he thought bacteria was ruining his grain but it wasnt showing on agar or LC. he was alluding to the grain having some byproduct that prevents grain from colonizing perhaps from fermentation or extremely dirty grain.
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SirPsycho
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SQuote:
LadysKnight said:
https://www.shroomery.org/forums/showflat.php/Number/24850162/fpart/1/vc/1
No, it doesn't grow into the agar. The food coloring being eaten away certainly doesn't show that. Mycelium release digestive enzymes into the environment, those breakdown food sources which the mycelium absorbs. That's what's going on.
Also if it was growing through the agar it would come out through the other side but it doesn't.
Yes, it will grow onto the wood of slants if it's exposed and most of the times it is because it floats in the solution so the top is almost always exposed.
It certainly doesn't grow into the agar. You can see it re-colonize when taking transfers from slants. I use a loop to cut out strips. They get colonized from the sides where myc is still present to the middle. If the myc were throughout the agar it would just grow out of the exposed layer and colonize the whole exposed surface at once.
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Mycolorado
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Re: No-pour agar sucks! [Re: SirPsycho] 1
#28101057 - 12/16/22 07:57 PM (1 year, 3 months ago) |
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This pretty clearly growing into the agar.
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SirPsycho
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Quote:
Mycolorado said:
This pretty clearly growing into the agar.
Where exactly is it supposed to be clearly growing into the agar?
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Mycolorado
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Re: No-pour agar sucks! [Re: SirPsycho]
#28101064 - 12/16/22 08:01 PM (1 year, 3 months ago) |
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SirPsycho
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Quote:
Mycolorado said:
Yep, that's a nice rhizo. How is it clearly in the agar?
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Mycolorado
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Re: No-pour agar sucks! [Re: SirPsycho]
#28101071 - 12/16/22 08:03 PM (1 year, 3 months ago) |
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Thatβs the bottom of the plate.
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SirPsycho
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Quote:
Mycolorado said:
Thatβs the bottom of the plate.
I understand that
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B Traven
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Re: No-pour agar sucks! [Re: SirPsycho] 1
#28101075 - 12/16/22 08:07 PM (1 year, 3 months ago) |
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I'm not gonna lie, I couldn't get through the OP much less the rest of this thread, so I'm just leaving my .02 after a cursory skim....
I run pasty plates and pyrex petri dishes, all no-pour style. I literally don't even have an agar recipe. I throw some shit in a saucepan and boil it, eyeballing the ratios and then boiling off or adding water to get the desired consistency. That means my agar varies widely in its basic composition and water content. I let the agar cool and solidify on the countertop before putting lids on, also wipe off any visible condensation. I wrap my plates in foil, usually quite well but sometimes more half-assedly if I'm re-using a bunch of janky torn pieces or whatever. Overall, I'm just not very meticulous about it.
Sometimes I get more condensation, sometimes less. Every once in a while I get plates flooded with water. Sometimes I don't use them, sometimes I use them anyway. Extra water and condensation in plates affects me about as much as rainshowers on a cross-country road trip. I barely notice or pay attention to them.
My more insidious contamination issues always trace back to specific cultures. The level of moisture present in some plate somewhere along the line never appears to be the deciding factor. If it were, then multiple culture lines would be fucked up by a few wet batches. But that's never what happens. It's always the same suspect lines kicking out bacterial spawn, and the same clean ones kicking out 
My more obvious contamination issues, I can confidently say, likely stem from my still-crappy SAB technique.
Doesn't matter how much excess moisture you have, you still need a source of contamination.
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Mycolorado
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Re: No-pour agar sucks! [Re: SirPsycho]
#28101078 - 12/16/22 08:14 PM (1 year, 3 months ago) |
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This the top.
Those rhizomes were growing off of the fruit. It was apparent that they were below the surface of the agar. 
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SirPsycho
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Quote:
Mycolorado said:
This the top.
Those rhizomes were growing off of the fruit. It was apparent that they were below the surface of the agar.
Seems to me like you can see the rhizome on the right dotted by the tomentose patches.
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ruawakeyet
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These rings on this PHV germ plate are also clearly growing into the agar, unless that isn't mycelium. Whatever it is, it grew into the medium. (I guess I should transfer a piece of one of them, duh!) (That "duh!" is to myself BTW.) It becomes much more evident in person when you can move the jar around and see the different angles.
This is an old batch of extremely weak GWA.
However, I don't see any relevance whatsoever to my searching for a reason why my no-pour will not readily contaminate. I really didn't want that debate either, I wish it wasn't even brought up again.
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