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Amazon Shop for: ½ Pint Jars, Agar, Malt Extract, Petri Dish, Potato Dextrose

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OfflineIGnosticAbhorI
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Agar's fun
    #4339019 - 06/26/05 01:36 PM (11 years, 5 months ago)

Put PDA mix in petri dishes.

PC for 20 mintues.

Then let cool for 12 er 24 hours.

Then knock up.

Isolate. Ect ect.

Sound about right so far?

Oh yea...Pretty sure this will work...lemme know what you think...I made a petri dish. :smirk:

1.Found old Altoids case.
2.Made a hole w/ screw driver.
3.Bent edges back.
4.ON the inside I covered the hole with transpore tape :smirk:
5.Outside I used a tiny bandaid...

Does that sound like it's all ready to go?

Already stuck it in the PC :smirk:

-Gnostic


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OfflineIGnosticAbhorI
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Re: Agar's fun [Re: IGnosticAbhorI]
    #4339047 - 06/26/05 01:45 PM (11 years, 5 months ago)

So how about I walked away to take a piss...

Took the weight off the PC, it whistled for about 3 mintues striaght.

NEVER WALK AWAY FROM A PC......mainly when you leave the burner on HIGH.

:doh:

damn that sucks...

-Gnostic


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OfflineIGnosticAbhorI
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Re: Agar's fun [Re: IGnosticAbhorI]
    #4339355 - 06/26/05 04:02 PM (11 years, 5 months ago)

Well...

Pc-ed it for the 20 minutes....and it's been sitting in the pc since.

PC's still warm to the touch so i'm sure the Agar is too.

Do I want to leave it in there or take it out to cool?

I've done grains plently of times...Didn't want the Agar to dry out - Being hot and getting cool to quick..

Er is it not like that?...:shrug:

tnkx

-Gnostic


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Offlinelepiota
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Re: Agar's fun [Re: IGnosticAbhorI]
    #4339401 - 06/26/05 04:28 PM (11 years, 5 months ago)

i also have made agar work today:
200ml hot tap water
4gr blended thai agar
6gr malt extract
all in a filtered jar (same kind of jar i use for LC etc...).
PCed for 20 mins.
open PC and take the jar out.
when agar jar is warm but not hot, agar is still liquid, then i inject through filter 2ml H2O2 and shake a little.
suck agar in a big syringe (60ml) and squirt in petris, 20ml in each petri.
when agar is solid and cold proceed with inoculation, in my case with squares of agar taken from previous generation.
picture: the 2 best dishes of the previous generation i used to inoculate this one.



hope this was useful! next time i'll make a pictorial if you think it is useful


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OfflineIGnosticAbhorI
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Re: Agar's fun [Re: lepiota]
    #4339412 - 06/26/05 04:33 PM (11 years, 5 months ago)

Ok...:thumbup:

So you use something like a pint jar or smaller...

PC...then use syringes - Put liquid in petries...

...Why not just pc the petri dishes?

I guess you could get more out of it if you did it that way.

But I don't need a bunch...I'm doing like 4-5..

Just used pints standing up with lids with water in em...

Placed petri on top and then pc-ed as normal...hmmm..

I don't want to try the jar way, mainly b/c when you do that whole part where you put the liquid in the petris...you increase contam rate.

-Sucking up liquid
-Placing in perti
...

Could just...skip that part :smirk:

But yea, make a pictorial, that'd own.

Thanks for the reply man.

:cheers:

-Gnostic


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Offlinelepiota
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Re: Agar's fun [Re: IGnosticAbhorI]
    #4339431 - 06/26/05 04:48 PM (11 years, 5 months ago)

>So you use something like a pint jar or smaller...

i use a small jar with filters like these:



they can hold 300ml, i put 200ml in it.

>...Why not just pc the petri dishes?

i sterilize petris in the oven, so they are dry when they come out. if i put them in the PC they are full of water when they come out, and taking out the water is incredibly messy.

if the syringe is sterile contam rate is kept to a minimum: the needle passes through a filter and sucks liquid. air is sucked into the jar when you do it, but sucked air passes through the other filter, so it is safe. moreover don't forget that i put h2o2 in agar.
agar is then injected in petris, so that i don't have to remove the petri lid (just lift a bit).

i will make pics next time i do it! :cool:


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OfflineIGnosticAbhorI
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Re: Agar's fun [Re: lepiota]
    #4339450 - 06/26/05 05:00 PM (11 years, 5 months ago)

Quote:

i will make pics next time i do it!




:headbanger:

Quote:

sterilize petris in the oven, so they are dry when they come out. if i put them in the PC they are full of water when they come out, and taking out the water is incredibly messy.




Hmmmm...Oven, lol...I wouldn't try that...

The pc...I thought that I'd have the same problem, but it worked out good.

If you place them higher than the water and higher than it'd boil up to...no prob...

But then again, I'm using Altoid cases :rotfl:...So I guess that they're a lil more snug than petris...:beer:

-Gnostic


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Offlinelepiota
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Re: Agar's fun [Re: IGnosticAbhorI]
    #4339494 - 06/26/05 05:15 PM (11 years, 5 months ago)

oven sterilization is not optimal, although i don't think that there are lots of bacteria or mold spores left in them after an alcohol wash and 20 mins at 300?F...
i'm having about 80% success rate with this method, and i think that the wrong 20% comes from somewhere else, i probably touch too much during inspections.


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OfflineIGnosticAbhorI
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Re: Agar's fun [Re: lepiota]
    #4339522 - 06/26/05 05:22 PM (11 years, 5 months ago)

What kinds of contams do you get?

The 20%...:smirk:

It is typically, green,cob...ect ect

I was thinking if it was green then maybe you might want to crank the oven up to 400F+...:grin:

Either that or it's somewhere along when you suck up the Agar (syringes)...:shrug:

-Gnostic


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Offlinelepiota
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Re: Agar's fun [Re: IGnosticAbhorI]
    #4339556 - 06/26/05 05:35 PM (11 years, 5 months ago)

the 2nd generation was plagued by some strange contam, maybe bacteria (i still don't know what it was), see that thing growing from the top left end of the petri:



3rd generation was almost ok, but while working i noticed that the worst growing dish also had a strange smell, probably another bacterium contam, this time it was invisible (first petri on the left of the top row).



another petri also smelled strange, but i'm not sure whether it was a contam or not. maybe only dog food smell, since the agar was DFA.


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Re: Agar's fun [Re: lepiota]
    #4342160 - 06/27/05 09:20 AM (11 years, 5 months ago)

Wow...It jellied up, lol.

Knocking it up today...Not sure what I should use...

Help me out here - Gimme some opinions.

-O/I Colonized Straw
-K.S.S multispore
-Creeper multispore
-Stropharia multispore


When isolating a strain, it doesn't matter if you use colonized sub. or multispore does it?

I don't think that it does, but anyways, gimme some opinions!

Tc

-Gnostic


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OfflineArcanePerception
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Re: Agar's fun [Re: IGnosticAbhorI]
    #4342307 - 06/27/05 10:59 AM (11 years, 5 months ago)

how do you see the myc growth or contaims through the altoid case?


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OfflineIGnosticAbhorI
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Re: Agar's fun [Re: ArcanePerception]
    #4342335 - 06/27/05 11:11 AM (11 years, 5 months ago)

:rolleyes:

Don't....lol

Acually, Next time I might put some type of plastic on first..then the lid...That way I can knock up, Take the lid off, and then see right through it.

But, yea, I can't, lol.

But I don't have petri dishes and can't really get em right now, SO, this way will suffice for now.

-Gnostic


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Offlinelepiota
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Re: Agar's fun [Re: ArcanePerception]
    #4342345 - 06/27/05 11:13 AM (11 years, 5 months ago)

i started this experiment with spores, i scraped spores directly on the agar. you can do it also by cloning (a piece of young shroom), a spore syringe, a LC, a colonized grain etc... everything. don't worry if the first generation will look ugly, you will have perfectly isolated strains only at later generations. my first generation looked like this:



i isolated the fastest and whitest piece for the second generation and so on...


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OfflineIGnosticAbhorI
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Re: Agar's fun [Re: IGnosticAbhorI]
    #4342373 - 06/27/05 11:21 AM (11 years, 5 months ago)

Wow, that is some ugly growth :roftl:

I just wanted someone to pick which strain....lol...

I think i'll do more work with creeper :grin:..

Tnkx guys, i'll probably update later...

-Gnostic


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Amazon Shop for: ½ Pint Jars, Agar, Malt Extract, Petri Dish, Potato Dextrose

Mushrooms, Mycology and Psychedelics >> Mushroom Cultivation

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