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InvisibleKatuXiK
Male

Registered: 09/19/13
Posts: 175
2nd agar attempt
    #18977500 - 10/14/13 05:02 PM (10 years, 3 months ago)

Just started my second agar attempt, using a difference recipe this time.

100ml tap water
1/4Tbsp agar
1/3tsp potato flakes
1/4tsp honey
small portion of tetracycline

Steam sterilized for about 20 minutes, didn't want to risk caramelizing again.

Just wondering, about how long does it take for agar to harden? It's been about an hour and a half at this point, and there is still a little bit on condensation in a couple of the dishes. They appear to be hardened nicely right now, but I figured I'd ask what you guys think.


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InvisibleStool_Sample
Fruity
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Registered: 09/25/12
Posts: 223
Loc: North East, US
Re: 2nd agar attempt [Re: KatuXiK]
    #18977540 - 10/14/13 05:14 PM (10 years, 3 months ago)

Not sure if you poured or made directly into petri's. It usually take my agar two hours to cool enough to hold with a mechanix glove, in order to pour into petri dishes.
Then I make sure to stack all petri's and then put the empty hot bottle ontop of the stack and leave to cool. By putting the hot bottle on top, it helps with condensation.
I usually only get One sometimes Two dishes with condensation.
then seal with parafilm and wait four days to see condition of petri's.

Hope some of that helped, good luck.


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InvisibleKatuXiK
Male

Registered: 09/19/13
Posts: 175
Re: 2nd agar attempt [Re: KatuXiK]
    #18977583 - 10/14/13 05:24 PM (10 years, 3 months ago)

Well, I sterilized and immediately after poured into my dishes, stacked them as 5 and let them cool. In an hour and a half they were cool to the touch and look completely solidified, there is no movement at all with tilting. I just inoculated 1 section in 4 dishes, and left 1 dish with all 3 sections uninoculated.


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InvisibleStool_Sample
Fruity
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Registered: 09/25/12
Posts: 223
Loc: North East, US
Re: 2nd agar attempt [Re: KatuXiK]
    #18978536 - 10/14/13 08:35 PM (10 years, 3 months ago)

Sounds like you jumped the gun on inoculation.
IMO you should give your Agar at least 48 hours to ensure no contams.

Also, not sure if you had condensation issues but if do less stacks, you will get less condensation.
(Just in case you had that issue)

Again, Good luck on the cultures.


--------------------
"wise words here"
:toadkart:


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InvisibleKatuXiK
Male

Registered: 09/19/13
Posts: 175
Re: 2nd agar attempt [Re: Stool_Sample]
    #18978649 - 10/14/13 08:54 PM (10 years, 3 months ago)

I didn't have any condensation that was enough to drip on the cake, just small droplets (like pin point size) on the lid, nothing on the agar. If I did jump the gun it's not a big deal, my supply of agar and potato flakes is limitless, the petri's require a trip to the surplus store, but that's not a big ordeal.

I might run out of spores pretty soon, but my first round of cakes is fruiting right now, so cloning is not far off. :smile:


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InvisibleKatuXiK
Male

Registered: 09/19/13
Posts: 175
Re: 2nd agar attempt [Re: KatuXiK]
    #18979606 - 10/15/13 12:47 AM (10 years, 3 months ago)

Hey I just realized, does my recipe sound okay or is that way off? Nobody commented on it and I didn't think to ask again. I just threw a pin the broke off on the agar, as far as I'm aware that's how cloning works... I also took of the string of mycelium it was attached to, and placed that on a tray as well. All in all I wound up with 2/4 of a pin in 2 trays, and mycelium in another 2 trays. Didn't initially plan this, but I was worried the pin might rot. I ate the other half of the pin cause I didn't know what shrooms taste like, I don't see why people say they taste so bad, tastes like it smells.


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OfflinePussyFart
Retired Cultivation Extrodinaire
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Registered: 04/08/12
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Re: 2nd agar attempt [Re: KatuXiK]
    #18979617 - 10/15/13 12:51 AM (10 years, 3 months ago)

You don't need honey or tetracycline....just agar and potatoe flakes is fine.

I just use tap water, agar powder, and light malt extract.

20g of each per liter of water.

KISS = Keep it simple stupid.


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InvisibleKatuXiK
Male

Registered: 09/19/13
Posts: 175
Re: 2nd agar attempt [Re: PussyFart]
    #18979646 - 10/15/13 12:58 AM (10 years, 3 months ago)

Wait you don't honey or anything? What is the point of all the crazy dextrose or LME methods? If I had know that I wouldn't have used honey, it's got so much bacteria! Is my sterile method alright, and did I do the right thing with the pin and mycelium? I'm pretty sure that is what I've read to do, but I've done stupid things in the past, figure I should probably ask.


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OfflineKizzle
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Re: 2nd agar attempt [Re: KatuXiK]
    #18979726 - 10/15/13 01:21 AM (10 years, 3 months ago)

Quote:

What is the point of all the crazy dextrose or LME methods?




The potato dextrose is the source of nutrients and the agar agar is for consistency, so you can pour it and get a nice smooth surface. There's no need to add anything else.

Same with the malt extract. As far as I know PDA and MEA work equally well for mushroom cultures.


Edited by Kizzle (10/15/13 01:27 AM)


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InvisibleKatuXiK
Male

Registered: 09/19/13
Posts: 175
Re: 2nd agar attempt [Re: Kizzle]
    #18979749 - 10/15/13 01:32 AM (10 years, 3 months ago)

Huh, was I reading outdated info? Or is there some minor benefit to each method?


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OfflineKizzle
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Re: 2nd agar attempt [Re: KatuXiK]
    #18979769 - 10/15/13 01:39 AM (10 years, 3 months ago)

Tetracycline is for making antibiotic agar but I have no clue why anyone would add honey :lol:


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InvisibleKatuXiK
Male

Registered: 09/19/13
Posts: 175
Re: 2nd agar attempt [Re: Kizzle]
    #18980263 - 10/15/13 07:28 AM (10 years, 3 months ago)

Because that's what the recipe I originally used called for. Agar, potato flakes, and honey. That recipe wound up not really working out for me, so I changed the agar and water amounts, but kept the honey and potatos the same. If I had known that I didn't need a sugar medium, I would've skipped it. I thought I did a pretty good amount of reading on this too..


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InvisibleSpitballJedi
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Registered: 10/13/12
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Re: 2nd agar attempt [Re: KatuXiK]
    #18980341 - 10/15/13 07:58 AM (10 years, 3 months ago)

potatoes are high in starch. starch is made of glucose. glucose is a sugar. Potato agar is a sugar media.


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InvisibleKatuXiK
Male

Registered: 09/19/13
Posts: 175
Re: 2nd agar attempt [Re: SpitballJedi]
    #18980788 - 10/15/13 10:36 AM (10 years, 3 months ago)

It's amazing the things you don't realize you've forgotten, a year ago I would've known that right off the bat; I can't believe I didn't make that connection... Thanks Jedi! :P


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InvisibleKatuXiK
Male

Registered: 09/19/13
Posts: 175
Re: 2nd agar attempt [Re: KatuXiK]
    #18986047 - 10/16/13 12:57 PM (10 years, 3 months ago)

No myc from MS yet, but both of my tissue drops and both of my myc drops are showing growth! Fuck yeah, science!


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