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Re: AGAR ENVY! (Anything and All things agar!) [Re: OldManRiver]
    #28653072 -

OldManRiver said:
AspectOfTheCreator said:
My t1 plates grow out clean. Is it risky or bad practice to use these for inoculating grain rather than going to t2 first?



I'll use them without concern.  I know that my transfer process is clean.  If you are confident that your transfers are reliable, I wouldn't hesitate. 

If the original plate was gross, I'd do a T2.



Thanks for reply. My germ plates hardly ever have contam on them anymore. The t1s always grow out perfectly symetrical and nice. Otherwise I wouldnt use them. I streak a Z on the germ plate and let it grow away from the streak mark before taking a t1 transfer. I scope the area out with a magbifying glass first too. The tiny little high powered area too.


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Re: AGAR ENVY! (Anything and All things agar!) [Re: AspectOfTheCreator]
    #28653471 -

Check out the aerial rope a dope going on right here

Storm Troopers T2. Really happy with the way it decided to do it's thing, & it's so fresh, & so clean :sunny:
:etjesus:


--------------------
"I'm a new Christ Savior - Neo from the Matrix
Young God negu, can't fuh with no lame shih
High off psychs, yeah, a negu stay t.a.d.e.d - Philanthropist - The Underachievers

T.A.D.E.D - Trippy All Day Every Day

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Re: AGAR ENVY! (Anything and All things agar!) [Re: MasterNeo]
    #28653492 -

MasterNeo said:
Check out the aerial rope a dope going on right here

Storm Troopers T2. Really happy with the way it decided to do it's thing, & it's so fresh, & so clean :sunny:
:etjesus:



That's purty.


--------------------
Sadly remembering when the Shroomery wasn't a closed minded mutual admiration society for OCD cargo cultists.  RIP methodical science.

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Re: AGAR ENVY! (Anything and All things agar!) [Re: OldManRiver] * 1
    #28653552 -

No go on the first attempt at Rancho Agosto germination but got some cool looking orange bacteria

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Re: AGAR ENVY! (Anything and All things agar!) [Re: SwabMarley]
    #28653601 -

How long has it been?

That's some slow looking bacteria. Could still see germination and be able to grab some if it hasn't been weeks and weeks.


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

culture storage

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Re: AGAR ENVY! (Anything and All things agar!) [Re: Nichrome]
    #28653606 -

Been 17 days so far. Yeah it’s very slow. I think this appeared around day 6ish. I will hold on to it for another week or so. If not I think there’s juuuust enough left on the print to have another go.

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Re: AGAR ENVY! (Anything and All things agar!) [Re: Tormato]
    #28653618 -

Hi all.

First time making agar plates with yeast: 2% agar, 1.6% LME, 0.2% nutritional yeast.  Same process followed for all my other agar - slowly vent PC, sterilise for 20 minutes, let cool down and pour at 50 degrees celsius.  Besides the plates looking butt ugly, today (day 4), my plates look some variation of this:



That looks very suspicious to me :rolleyes:  Is this what yeast plates are supposed to look like?

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Re: AGAR ENVY! (Anything and All things agar!) [Re: SWBZA]
    #28653646 -

SWBZA said:
Hi all.

First time making agar plates with yeast: 2% agar, 1.6% LME, 0.2% nutritional yeast.  Same process followed for all my other agar - slowly vent PC, sterilise for 20 minutes, let cool down and pour at 50 degrees celsius.  Besides the plates looking butt ugly, today (day 4), my plates look some variation of this:



That looks very suspicious to me :rolleyes:  Is this what yeast plates are supposed to look like?



Looks suspicious to me too. Could be pouring/sterile technique, but I think 20 minutes may be too short to properly sterilize the amount of media you used, especially with more particulate than you may usually have. See e.g. https://www.shroomery.org/forums/showflat.php/Number/28166056

Edited by Phaethon (02/10/24 02:41 AM)

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Re: AGAR ENVY! (Anything and All things agar!) [Re: Phaethon]
    #28653649 -

I go up to an hour on liquid culture.
I go like 50 to 55 minutes on Petri dishes too.

I just cover the jars with foil, so water doesn’t get into the filters.
But why not?

How long is too long to sterilize?

I feel like I shouldn’t have to go past an hour.

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Re: AGAR ENVY! (Anything and All things agar!) [Re: Phaethon]
    #28653678 -

Interesting post by SPG. Thanks for the pointer. I'll give my next batch a longer cook. I took the time for a recommendation to balance sterility against caramelization for LCs. I'm going to have to go back to my notes - getting confused working from memory, I think. Okay, suckers, next time I'm sterilising the living daylights out of you, little bugs! Anyway, don't think I'll do yeast plates again anytime soon. They look ugly!

Edit: just checked the SPG formula: 32 minutes for me.  So plates probably not all clean.  Bummer.

Edited by SWBZA (02/10/24 04:28 AM)

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Re: AGAR ENVY! (Anything and All things agar!) [Re: SWBZA]
    #28653815 -

here’s some T2 plates. What do you think is up with the denser ring of mycelium that all of the plates have near the outer edge? Do these look healthy? :smile:


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spinning, spinning, spinning... spun
💃🏻💀⚡️

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Re: AGAR ENVY! (Anything and All things agar!) [Re: scwoopz]
    #28653855 -

scwoopz said:
here’s some T2 plates. What do you think is up with the denser ring of mycelium that all of the plates have near the outer edge? Do these look healthy? :smile:



I'm also curious to see the answer - I've had rings before as well.  There was a post I read where the person said something like "you can see the temperature changes of the last days" when describing such rings.  I can't remember the post details.  Did you have a temperature change?

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Re: AGAR ENVY! (Anything and All things agar!) [Re: SWBZA]
    #28653860 -

SWBZA said:
Interesting post by SPG. Thanks for the pointer. I'll give my next batch a longer cook. I took the time for a recommendation to balance sterility against caramelization for LCs. I'm going to have to go back to my notes - getting confused working from memory, I think. Okay, suckers, next time I'm sterilising the living daylights out of you, little bugs! Anyway, don't think I'll do yeast plates again anytime soon. They look ugly!

Edit: just checked the SPG formula: 32 minutes for me.  So plates probably not all clean.  Bummer.



Now we wait and see.  Hopefully I'll be able to declare clean agar in about a week.


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Re: AGAR ENVY! (Anything and All things agar!) [Re: scwoopz]
    #28653891 -

scwoopz said:
here’s some T2 plates. What do you think is up with the denser ring of mycelium that all of the plates have near the outer edge? Do these look healthy? :smile:



Looking great. Those are normal and the result of natural growth cycles. Think rings on a tree.


--------------------
A Love Letter to New Growers
The 7 Edicts of Mushroom Cultivation as Decreed by Smellyhobbit

:kodama:


Anyone can get a canopy. But can you get what you want?

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Re: AGAR ENVY! (Anything and All things agar!) [Re: SwabMarley] * 1
    #28653895 -

SwabMarley said:
Been 17 days so far. Yeah it’s very slow. I think this appeared around day 6ish. I will hold on to it for another week or so. If not I think there’s juuuust enough left on the print to have another go.



If I get bacteria on germ plates, I just transfer to brf puck (if the spores germinated) then the bacteria disappears. Love that brf shit lol. But you have more spores so eh

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Re: AGAR ENVY! (Anything and All things agar!) [Re: SWBZA] * 1
    #28654092 -

SWBZA said:
Hi all.

First time making agar plates with yeast: 2% agar, 1.6% LME, 0.2% nutritional yeast.  Same process followed for all my other agar - slowly vent PC, sterilise for 20 minutes, let cool down and pour at 50 degrees celsius.  Besides the plates looking butt ugly, today (day 4), my plates look some variation of this:



That looks very suspicious to me :rolleyes:  Is this what yeast plates are supposed to look like?




That is bacteria. Toss all plates that look like that right in the trash. Some kind of flaw in your method.


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

culture storage

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Re: AGAR ENVY! (Anything and All things agar!) [Re: GenesisCorrupted]
    #28654097 -

GenesisCorrupted said:
I go up to an hour on liquid culture.
I go like 50 to 55 minutes on Petri dishes too.

I just cover the jars with foil, so water doesn’t get into the filters.
But why not?

How long is too long to sterilize?

I feel like I shouldn’t have to go past an hour.




For LC without solids I do 15 minutes at 15 psi post venting. For anything with solids in the liquid like BRF LC I do 30 minutes.

Agar, 45 minutes but that may be overkill.

With sugar based broths you start to see more caramelization after about the 40 minute mark. If I forget to set a timer and they go long like that the broth is darker and doesn't preform as well. Timing will depend on ingredients for example fructose doesn't seem to brown as easily as malt or rice syrup.


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

culture storage

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Re: AGAR ENVY! (Anything and All things agar!) [Re: Nichrome]
    #28654103 -

I use telephone brand, agar with potato flake and some food coloring.

But I see no reason why I shouldn’t attempt less time and see if it affects the result.

My stuff seems to like a stiff agar.

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Re: AGAR ENVY! (Anything and All things agar!) [Re: GenesisCorrupted] * 1
    #28654111 -

45 at 15 post vent should be fine.


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

culture storage

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Re: AGAR ENVY! (Anything and All things agar!) [Re: Nichrome]
    #28654121 -

Nichrome said:
SWBZA said:
Hi all.

First time making agar plates with yeast: 2% agar, 1.6% LME, 0.2% nutritional yeast.  Same process followed for all my other agar - slowly vent PC, sterilise for 20 minutes, let cool down and pour at 50 degrees celsius.  Besides the plates looking butt ugly, today (day 4), my plates look some variation of this:



That looks very suspicious to me :rolleyes:  Is this what yeast plates are supposed to look like?




That is bacteria. Toss all plates that look like that right in the trash. Some kind of flaw in your method.



Thanks, they are gone. Ouch, time to go back to basics.

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