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OfflineFlusH
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Agar help please
    #25832214 - 02/23/19 02:25 PM (2 years, 10 days ago)

so I've been following pastywhites no pour agar tec as closely and completely as possible.

in the pictures, it's hard to see but there's no growth at all on the agar. the jars in the background were inoculated at the same time with the same syringe.



any ideas why the agar is either slower or not working?


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OfflineFlusH
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Re: Agar help please [Re: FlusH]
    #25832234 - 02/23/19 02:33 PM (2 years, 10 days ago)

jars/agar was inoculated Jan 18 2019. pretty much just over a month ago.

strain is GT, used spore syringe.

this is my third attempt at agar without success yet.

any other info I can offer?


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Invisiblerm1024
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Re: Agar help please [Re: FlusH]
    #25832244 - 02/23/19 02:37 PM (2 years, 10 days ago)

Try to get clearer pictures.  How much solution did you use per plate?  You will have much better luck at getting help if you can get better pics.

cheers


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OfflineFlusH
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Re: Agar help please [Re: rm1024]
    #25832448 - 02/23/19 03:56 PM (2 years, 10 days ago)

I absolutely agree. I have a pixel 2 unfortunately I dropped it several times 😂

I'll get better pictures in a couple hours


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OfflineFlusH
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Re: Agar help please [Re: FlusH]
    #25832826 - 02/23/19 06:48 PM (2 years, 9 days ago)



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OfflineFlusH
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Re: Agar help please [Re: FlusH]
    #25832836 - 02/23/19 06:51 PM (2 years, 9 days ago)

k, so I got frustrated trying to get a decent picture, and ended up opening up two just to get a better look.

when I dropped the spores from the syringe, there was a noticable droplet with visible spores. can't see them in the pics above.

the other non green containers are recent ones I just made three days ago, half with syringe and half with loop.


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Offlinevikingsc
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Re: Agar help please [Re: FlusH] * 2
    #25832981 - 02/23/19 07:33 PM (2 years, 9 days ago)

What's the exact recipe you're using to make your agar? Perhaps not nutritious enough?

Your doing PF Tek so you obviously have BRF. Try making BRF agar?
2% Agar and 1.5% - 2% BRF for germinating spores
and 0.5% - 1.5% BRF for transfers and growing out.

So for germinating spores at 2% BRF for every 100ml of water add 2 gr of agar powder and 2 gr of BRF and whatever food coloring you want to add.


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OfflineFlusH
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Re: Agar help please [Re: vikingsc]
    #25833706 - 02/24/19 01:18 AM (2 years, 9 days ago)

This is taken directly from Pastywhyte's Easy Agar tek, and is what I am following

Quote:

I then spoon 2 heaping tablespoons into the pot. I rarely weigh it out, if I want stiffer agar I use a little more, if I want softer I use a little less. If you want to weigh it, for this amount of agar you will want about 2-2.5 grams depending on how soft or firm you want it to be. If your using powdered agar and don't have a scale, a level tsp (5ml) is roughly 2.5 grams. Remember that no matter how much you want to mix up, you want a ratio of 20-25 grams of agar to 1 liter of water. If you have issues with the mix being too stiff while mixing then adjust the ratios for less agar. Some food grade agars will work out to be a little stiffer than others, some experimentation may be required.

If all that is a bit confusing the just remember 2% is roughly what you are aiming for.

Add 2 teaspoons (10 ml) of potato flakes, and 1/2 cup (120 ml) of water (tap is fine). Then I turn the heat on low on the stove a stir the mixture until its mostly dissolved. At this point I will add a drop of Karo or Honey. Stir it all up well. It does not matter if there are still a few pieces of agar still not dissolved, just get most of it. I like to use either colored agar (some places have this) or else just add a few drops of food coloring to the mix. This will help you see growth real quick, and the contams are easily spotted as well. There will be nowhere for them to hide :lol: If you can only get regular agar, I like to use green food coloring to make the contams stand out best, but any dark color (red, blue, purple) will work as well :thumbup:



https://www.shroomery.org/forums/showflat.php/Number/24357122#24357122


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OfflineFlusH
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Re: Agar help please [Re: FlusH]
    #25833714 - 02/24/19 01:24 AM (2 years, 9 days ago)

I will give the BRF agar mix a shot though,  Cant hurt at this point. Also Im thinking of trying to clone on to agar, and hope that has better results.


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InvisibleBigSurMoon
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Re: Agar help please [Re: FlusH] * 1
    #25834047 - 02/24/19 09:31 AM (2 years, 9 days ago)

Did you spread the spores around? Or just a drop? Cause if you just put a drop, there's a good chance it just rolled around on the agar while handling it. Spores on agar do take a while to show up, I have had it take two weeks before showing anything. Fyi, brf is a pretty thick agar, I suggest filtering it once or so, Ive done brf a couple times, and I didn't really like it, but that doesnt mean it's not good though, lol.


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I swear that "black kow experiment" thread is gonna get it. It's tempting me to adress it. Sounds both pornografic and racist...
https://www.shroomery.org/forums/showflat.php/Number/25300352/page/1

REVERSE BIOPSY LC, (my twist on Josex's lc tek.)
https://www.shroomery.org/forums/showflat.php/Number/25986733/ center]

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OfflineApc123
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Re: Agar help please [Re: BigSurMoon] * 1
    #25834280 - 02/24/19 11:41 AM (2 years, 9 days ago)

My favorite agar recipe by far is - 10g agar to 12-15g light malt extract to 500ml water - so simple and easy. Comes out perfect every time. (food coloring optional) :super:


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Invisiblerm1024
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Re: Agar help please [Re: Apc123] * 1
    #25834371 - 02/24/19 12:18 PM (2 years, 9 days ago)

Those plates unfortunately aren't looking very good from where I am sitting, but I have been wrong before.  I can't speak much to your agar recipe as I am more familiar grainwater agar.  I see much more resilient growth from the grainwater. 
On your next go you might try getting some sterile swabs and dropping a single droplet of spore solution onto that and then doing a proper streak on the plate.
Talk a little bit about your pressure cooking procedures as that could be an area where you might be having trouble.
Regarding your what appear to be PF jars, they don't look so great to me but Im not PF expert so someone else might chime in here.
hth


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Re: Agar help please [Re: rm1024] * 1
    #25834401 - 02/24/19 12:26 PM (2 years, 9 days ago)

I followed pastys recipe and had zero results with it in my experience, but i figured that was because i was using actual agar powder and not the stuff he was using. I adjusted the PDA to have less potato and more water for my powder and have had great results. Just fiddle with it using some test myc from your jar to see how well it grows with the varying recipes i found the more jelly and less solid the agar the better results i had when germinating on the plates.

I started with this recipe and fiddled with it till i got it to how i like, You should do your own experimenting to find the values you prefer.
Quote:

7.5 grams of dried potato flakes.    | 2 tsp
13.5 grams of powdered agar agar. | 1 tsp
10.5 grams of dextrose.                      | 1 drop
750ml of distilled water.                    | 120ml





That being said im definitely switching over to grainwater agar for some clearer views through the plate since the potato really fogs it up.


Edited by drudigger (02/24/19 12:29 PM)


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OfflineFlusH
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Re: Agar help please [Re: drudigger]
    #25835507 - 02/24/19 10:37 PM (2 years, 8 days ago)

Quote:

drudigger said:
I followed pastys recipe and had zero results with it in my experience, but i figured that was because i was using actual agar powder and not the stuff he was using. I adjusted the PDA to have less potato and more water for my powder and have had great results. Just fiddle with it using some test myc from your jar to see how well it grows with the varying recipes i found the more jelly and less solid the agar the better results i had when germinating on the plates.

I started with this recipe and fiddled with it till i got it to how i like, You should do your own experimenting to find the values you prefer.
Quote:

7.5 grams of dried potato flakes.    | 2 tsp
13.5 grams of powdered agar agar. | 1 tsp
10.5 grams of dextrose.                      | 1 drop
750ml of distilled water.                    | 120ml





That being said im definitely switching over to grainwater agar for some clearer views through the plate since the potato really fogs it up.





never thought of it till you mentioned it. I'm using agar powder as well.

thanks for that, I'll try your recommendations


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OfflineFlusH
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Re: Agar help please [Re: Apc123]
    #25840835 - 02/27/19 11:16 AM (2 years, 6 days ago)

Quote:

Apc123 said:
My favorite agar recipe by far is - 10g agar to 12-15g light malt extract to 500ml water - so simple and easy. Comes out perfect every time. (food coloring optional) :super:





so I tried this.. and got this
😂😂😂🤣🤣

I'm guessing 10g agar wasn't enough for me lmao.. back to trying again


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Invisiblenube424
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Re: Agar help please [Re: FlusH]
    #25840902 - 02/27/19 11:51 AM (2 years, 6 days ago)

What type of agar? And are u SURE ur measurements were right? 🙄 2% agar comes out pretty stiff. Even the cheap Asian store bar agar.


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OfflineFlusH
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Re: Agar help please [Re: nube424]
    #25841127 - 02/27/19 01:21 PM (2 years, 6 days ago)

I see what you're saying and I've been doing some investigating..

I'm using agar from a local health food store. they must buy it in bulk and repackage it and place their own stickers on it.  this agar that I have tastes incredibly sweet by itself.

I went and bought a telephone name brand from the city a few miles away.  no Sweet taste..

I'm thinking this health food store has been cutting their pre package agar with dextrose 🤔


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Invisiblenube424
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Re: Agar help please [Re: FlusH]
    #25841179 - 02/27/19 01:49 PM (2 years, 6 days ago)

Anything's possible in retail lol. I get the bars from the Asian grocery stores. It works great but it has food coloring


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Mushrooms, Mycology and Psychedelics >> Mushroom Cultivation

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