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Mushrooms, Mycology and Psychedelics > Mushroom Cultivation
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Re: AGAR ENVY! (Anything and All things agar!) [Re: D3_Myc]
    #26846835 -

Having some issues with no-pour plates becoming contaminated before even inoculation.  I made some(25) no-pour plates about a week ago.  I boiled the agar/lme/water, poured them into the prepared containers and then PC'd them @12psi for 2 hours.  I let them cool a bit, removed them from the PC once depressurized and let them sit until today. Only 7/25 were contam free, before even opening them!  I did NOT use glad mini-rounds.  I used another pp5 container/lid and only 1 layer of tape.  Should I use mini-rounds and several layers of tape?? Thanks for the advice!

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Re: AGAR ENVY! (Anything and All things agar!) [Re: D3_Myc]
    #26846856 -

D3monic said:
9, 11 and 3 is where I’d grab from



Thanks bud :cheers:

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Re: AGAR ENVY! (Anything and All things agar!) [Re: wolfXsoul]
    #26846955 -

I use 2-3 layers of micropore tape on my no pour stuff. Are you layering paper towels on top before covering with foil? I'm not sure if it's absolutely necessary, but I always do it because I worry that a wet filter will let in contaminants. It's what pasty recommends in his easy agar tek iirc.

Another unrelated thing I'll add is that you probably don't have to pc plates that long even at 12 psi. Definitely do a search and see what it works out to for 12 psi instead of the standard 15.


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Re: AGAR ENVY! (Anything and All things agar!) [Re: Tight Lunchbox]
    #26846992 -

So I made my first agar plates last week but did not inoculate. Week later I returned to my SAB, where I left them, to check on my BRF cakes and some bags of uncle Ben's I loaded up with spores and realized that I never taped them I just stuck them in a bag and left.
So now I want to inoculate the agar with some spore swabs and I'm wondering do I pressure cook the plates again ( cheap plastic plates used h2o2 at 6ml of 3% per liter ) also do I need to drill holes in lids? [url=https://files.shroomery.org/files/20-30/582784596-15958278099663750934160903327154.jpg

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Re: AGAR ENVY! (Anything and All things agar!) [Re: Tight Lunchbox]
    #26847275 -

Tight Lunchbox said:
I use 2-3 layers of micropore tape on my no pour stuff. Are you layering paper towels on top before covering with foil? I'm not sure if it's absolutely necessary, but I always do it because I worry that a wet filter will let in contaminants. It's what pasty recommends in his easy agar tek iirc.

Another unrelated thing I'll add is that you probably don't have to pc plates that long even at 12 psi. Definitely do a search and see what it works out to for 12 psi instead of the standard 15.


I'll check it out. 2 hours is what I do my Jars at, so I figured it was safe for agar too. I am putting a piece of paper towel on it, but it seems to stick to the tape and I'm wondering if it's damaging it.

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Re: AGAR ENVY! (Anything and All things agar!) [Re: wolfXsoul] * 1
    #26847296 -

I’d be concerned about caramelizing the agar after that long even at 12psi


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D3's Agar TEK
Agar Porn Gallery
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Re: AGAR ENVY! (Anything and All things agar!) [Re: wolfXsoul]
    #26847315 -

wolfXsoul said:
Tight Lunchbox said:
I use 2-3 layers of micropore tape on my no pour stuff. Are you layering paper towels on top before covering with foil? I'm not sure if it's absolutely necessary, but I always do it because I worry that a wet filter will let in contaminants. It's what pasty recommends in his easy agar tek iirc.

Another unrelated thing I'll add is that you probably don't have to pc plates that long even at 12 psi. Definitely do a search and see what it works out to for 12 psi instead of the standard 15.


I'll check it out. 2 hours is what I do my Jars at, so I figured it was safe for agar too. I am putting a piece of paper towel on it, but it seems to stick to the tape and I'm wondering if it's damaging it.


Poisoned's glass petri no-pour tek
Caps' SAGARI Method (Soft Agar Innoculant)
D3's Agar and Xfer TEK
c10's Agar Guide
Bod's Agar TEK
IMHE I have not had any success trying Pre-Pour, they come out a sloppy wet
mess and are more trouble than pouring in the SAB.


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:sporedrop: GLOSSARY  :sporedrop: ACROMYMS!   :sporedrop: GETTING STARTED :sporedrop:

Edited by Inthepit (07/27/20 10:08 AM)

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Re: AGAR ENVY! (Anything and All things agar!) [Re: Inthepit]
    #26847575 -

This is the 4th transfer away from spore on Texans. I've never seen a culture do this before. You think it's responding to a hidden contam or are the sectors just keeping distance from one another? In the second picture you can see where the gap had formed and then as it grew out the culture closed in a round it..




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Contam or myc? From a drop from multispore syringe [Re: Tormato]
    #26847609 -

Does this look like mycelium to you or bad stuff?  My first agar in over a year so feel very rusty at identifying.  Thanks. :smile:








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Re: Contam or myc? From a drop from multispore syringe [Re: senorpoto]
    #26847619 -

Trash them-- that ain't mecelium

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Re: Contam or myc? From a drop from multispore syringe [Re: senorpoto]
    #26847623 -

Looks suspect as heck.


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

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D3's Agar TEK
Agar Porn Gallery
RW x MelMak Grow Along Thread

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Re: Contam or myc? From a drop from multispore syringe [Re: senorpoto]
    #26847644 -

Thanks guys, will toss.

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Re: AGAR ENVY! (Anything and All things agar!) [Re: wolfXsoul]
    #26848239 -

I took tissue from a beautiful B+ cluster and dropped it on two different types of agar. One is premade MEA, the other is water from an oat boil + agar the way that Bod recommends. Interesting to see the different style of growth. I did several of these and they all turned out similar. I did notice that the Oat agar was much firmer in texture and seemed to be less optimal for spore germination when I tried that.

I'm also wondering about isolating sectors at this point. Do people that come up with amazing isolates usually do so from spores or a isolating from a clone. It's harder to see the sectoring on the oat agar, so if anyone could point out where they are that would be awesome.




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Re: AGAR ENVY! (Anything and All things agar!) [Re: beeker]
    #26849250 -

beeker said:
I took tissue from a beautiful B+ cluster and dropped it on two different types of agar. One is premade MEA, the other is water from an oat boil + agar the way that Bod recommends. Interesting to see the different style of growth. I did several of these and they all turned out similar. I did notice that the Oat agar was much firmer in texture and seemed to be less optimal for spore germination when I tried that.

I'm also wondering about isolating sectors at this point. Do people that come up with amazing isolates usually do so from spores or a isolating from a clone. It's harder to see the sectoring on the oat agar, so if anyone could point out where they are that would be awesome.






Your oat water agar is probably lower nute than the MEA, and the myc is hungry and looking for food. . . Tormentose (fluffy) growth is often a sign of well fed lazy myc. . .

Its hard to be consistent with grain water as you can't measure the amount of nute as you can with LME or any other sugar. . .

IMO much of the talk around isolates is educated speculation without microscopy verification. Take the leading end off one tendril off your oat plate, and transfer to a fresh plate. . . As this comes from a clone and is a clear tendril, you will be much closer to what is considerred an isolate.


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Re: AGAR ENVY! (Anything and All things agar!) [Re: Bobbit]
    #26849342 -

I've seen people rate Oat GW as the most potent nutrient wise GW. Take that for what you will. I'm seeing speckles of sediment, but the actual colour of the agar leads me to suggest its probably got enough nutes.


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Re: AGAR ENVY! (Anything and All things agar!) [Re: ManifoldPrime]
    #26849350 -

Next round of agar I run I planned on restricting nutes to maybe 60%-- I only see relatively fast uniform tomentose growth on everything-- I want that rhizo reaching out.

I already was using less than the standard LME ratio-- maybe 85% of the standard 10g per liter. Still tomentose only no matter what culture I've worked with so far.

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Re: AGAR ENVY! (Anything and All things agar!) [Re: coAsTal] * 1
    #26849413 -

I always use half strength malt for everything except germination plates.

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Re: AGAR ENVY! (Anything and All things agar!) [Re: ManifoldPrime]
    #26849894 -



Trying to clean up an ape swab. Any thoughts on wtf to do with this one as far as additional transfers and identifying contam?

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Re: AGAR ENVY! (Anything and All things agar!) [Re: ruK]
    #26850176 -

coAsTal said:
maybe 85% of the standard 10g per liter.


I think 20g is the standard per liter. Did you mean a half liter (500ml)?

ruK said:


Trying to clean up an ape swab. Any thoughts on wtf to do with this one as far as additional transfers and identifying contam?


I'm not an expert but I'm going to say toss it and start over. Not sure what contams you have (bacteria?) but I don't see anything worth transferring.

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Re: AGAR ENVY! (Anything and All things agar!) [Re: Sped105]
    #26850232 -

As far as i understand, standard from lab supply store MEA is 3%, or 30g per liter

Here people usually use half strength @15g per liter, which you could call mushcult standard

Although itll work pretty much regardless. I use less than 0.17 % (1.67g per L) in my LCs

Switching up recipe can give you different myc appearance,  but i like to keep it standardized so i can make comparisions without chalking up differences to the medium


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

C10's Agar Guide + Tips and Tricks | c10's Flow Hood Build Guide


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― Friedrich Nietzsche

Edited by c10h12n2o (07/29/20 02:50 AM)

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Mushrooms, Mycology and Psychedelics > Mushroom Cultivation
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