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Agar Poke-a-Plates


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#1 Sideshow

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Posted 25 August 2009 - 04:06 PM

I was really bored at work one day and came up with an alternative to petri dishes or mason jars for agar work. This is not a new tek, mind you - only a variation on threads posted on 'Topia. None of this would have been possible without Hip's airport and the Banzai Agar tek.
https://mycotopia.ne...s-banzai-style/

https://mycotopia.ne...c/3588-airport/
:bow:


I was looking for a container that is slightly smaller than the standard 150mm petri dish which was also disposable and easy to use. I found and liberated the following SOLO souffle cups:

DSCN0309.JPG

This is what the finished plate looks like:

In hand.JPG

They are very easy to use, store very well (about a year, more depending on how thick the agar is poured), and require no parafilm or other sealant.

So I start out by taking a bunch of new, empty cups into the glovebox and wiping them down inside and out with bleach water/alcohol/peroxide - Any one of those is fine. After wipng out each one, I put the caps on and remove a batch frome the glovebox. This time I used my brand spankin' new schmuvbox (thanks, eatyu)

Schmuv1.JPG
Schmuv2.JPG

I take my prepared agar from storage and heat it in a water bath.

Water Bath.JPG

And suck it into a big syringe. 10ml syringes can get you 2 plates if you are stingy.

To syringe.JPG

Then I wipe the lid with alcohol and inject the agar.

Alky wipe.JPG

Empty.JPG

Full.JPG

After I pull the syringe out, I tape over the hole with the smallest peice of scotch tape i can manage to pull off the roll

Taped 2.JPG

Bagged.JPG

I bag 'em up and throw them in a drawer for later use. As I said earlier, they are good for a minimum of 6 months when poured very thinly, about a year if I use 12ml or more of agar (the pint jar of prepared agar in the pics is about a year old, unrefrigerated, and I can't tell the difference between it and a batch made yesterday). I also like the fact that I can just shoot spores or lc right thru the 1st hole made in the lid. If anyone can tell me a good coring tek for cloning, I'd love to hear it because I can never get a good tissue sample from the shroom to the needle to the agar succesfully. My goal is to be able to bring these to FOAF's house, core a sample in open air, and then shoot it right thru the lid of these babies.

On a related note, I'm lookng for someone to point me to a supplier of luer lock needles larger than 16ga.
Thanks, and hope you enjoyed the thread!

Attached Thumbnails

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Edited by coorsmikey, 05 May 2016 - 08:58 PM.
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#2 Sideshow

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Posted 25 August 2009 - 04:09 PM

Oh and you can get the cups here:
http://www.wesellcof...com/page29.html

The ones I use are 2 oz, but some may like the 3.25oz better.
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#3 Elf Salvation

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Posted 25 August 2009 - 05:11 PM

trying this one out myself

:)
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#4 Foster

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Posted 25 August 2009 - 07:00 PM

Nice work man.:eusa_clap Hehe I happen to have a couple packs of similar containers from a well known ice cream /fast food restaurant.
Just might give this a try myself.

I have both black (sauce containers with lids), and clear with lids. I like that they seal better than petris. Now if they could be found in pp5 or pc'able material. Just be an added bonus.

Just knew there was a reason I kept those containers. Other than for shroomy jello shots.

Multi-use! I love that. Good thinking, even if it was mentioned before. Sometimes the re-thought, revised, reused, recycled info is the best.

Nice write up and pics also. Very easy to understand and replicate.

Thanks for sharing. :)

#5 roscoe

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Posted 25 August 2009 - 07:18 PM

i have no expirence with agar yet. but i read a clean work station is paramount. maybe not?


http://mycotopia.net...tes-syringe.jpg


love this pic!

#6 Ben Dover

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Posted 25 August 2009 - 07:36 PM

i have no expirence with agar yet. but i read a clean work station is paramount. maybe not?


Yeah, a sterile environment is important with agar use. You just don't want any bad germs landing on the agar when you are putting something on it.

That's what the 'schmuvbox' is for :)

#7 Sideshow

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Posted 25 August 2009 - 09:19 PM

Roscoe, you can definitely do agar in open air, ie the bathroom if you spray lysol or oust beforehand. The banzai agar tek is great cuz you only have to slightly lift the petri lid and squirt in agar from the syringe quickly - nothing to it. Believe me, agar can be done in some downright dirty conditions if the sterile technique is sound.

#8 roscoe

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Posted 26 August 2009 - 03:30 AM

Roscoe, you can definitely do agar in open air, ie the bathroom if you spray lysol or oust beforehand. The banzai agar tek is great cuz you only have to slightly lift the petri lid and squirt in agar from the syringe quickly - nothing to it. Believe me, agar can be done in some downright dirty conditions if the sterile technique is sound.


well if you can get it to work on the stove with a package of meat right next to it, then im sure i can make it work! gives all of us newbs hope. :bow:

#9 Sideshow

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Posted 28 August 2009 - 11:14 PM

3 days later, we have some myc growth from 2 lc's:

Myc1.JPG

Myc 2.JPG

A control (one of the 9 done before)

Control.JPG

And one that isn't looking well (the only one that has gone south)

Contam.JPG

#10 bozefonce

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Posted 28 August 2009 - 11:25 PM

After I pull the syringe out, I tape over the hole with the smallest peice of scotch tape i can manage to pull off the roll


Would a blob of silicone caulk on each lid be a better solution for multiple uses? It might work like the injection ports on BRF lids that I've seen elsewhere in the forum. The nice thing is that since these lids are thin plastic, you don't have to punch holes in the lids first or anything, just put a blob on and let dry. Then you can punch through that silicone many times before the lid becomes unusable. Cheap, easy, nice!
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#11 masterplain88

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Posted 28 August 2009 - 11:28 PM

what agar mix did you use. and you mentioned lc in one of the pics is it a lc mix in the cups.if not could a small amount of lc in a cup work.
masterplain88

#12 Sideshow

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Posted 28 August 2009 - 11:32 PM

what agar mix did you use. and you mentioned lc in one of the pics is it a lc mix in the cups.if not could a small amount of lc in a cup work.
masterplain88


It is MRYA or malt extract, brf, nutritional yeast & agar powder.
I used lc's on the agar to start the culture.
In reference to your lc question, this should do nicely:

http://mycotopia.net...tek-merged.html

#13 masterplain88

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Posted 28 August 2009 - 11:37 PM

yea i have seen that tek looks like it would work well now what are the bennefits of using agar compared to the lc method for strain isolation?
thanks,
masterplain88

#14 Sideshow

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Posted 28 August 2009 - 11:42 PM

2 advantages:
Agar is the only way to clean up a culture and
Agar allows more reliable substrain isolation than other methods, IMO

#15 Leftover Salmon

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Posted 29 August 2009 - 12:06 AM

2 advantages:
Agar is the only way to clean up a culture and
Agar allows more reliable substrain isolation than other methods, IMO


Now the way to achieve this is to use the best looking pieces of myc growth on the agar? I mean do you simply cut off the more aggressive looking myc and use this? Or do you isolate shit on a microscopic level...

Sorry just trying to wrap my head around this.

#16 weeeeeee

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Posted 29 August 2009 - 12:35 AM

i have no expirence with agar yet. but i read a clean work station is paramount. maybe not?


http://mycotopia.net...tes-syringe.jpg


love this pic!


thats why it looked like he was using the shmuvbox which from what i have heard is wonderful for agar

#17 Sideshow

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Posted 29 August 2009 - 08:06 AM

Now the way to achieve this is to use the best looking pieces of myc growth on the agar? I mean do you simply cut off the more aggressive looking myc and use this?


Exactly, just cut a square of agar that has the best looking myc on the plate- or you can srape some myc off the surface of the agar with a scalpel, etc and put it on a fresh one. Larger, traditional petri dishes are better for spore isolation because they are much larger and provde more surface area, giving the mycellium room to grow out and sector.

#18 BuckarooBanzai

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Posted 29 August 2009 - 10:47 AM

Nice Tek, man.

Archive Material - to Agar & Strain Isolation

Be careful storing agar plates for more than 5 or 6 months. Old agar can dry just a bit and then it just sucks. The agar looks the same but spores struggle to hatch and myc grows a lot more slowly.

Fresh plates work much, much better.

#19 Sideshow

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Posted 29 August 2009 - 11:33 AM

:bow:
Thanks, BB. you were the inspiration for all of this.

I have had really good luck using 8-12 month old plated agar, but maybe that's just me.

#20 Hippie3

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Posted 29 August 2009 - 01:32 PM

yea i have seen that tek looks like it would work well now what are the bennefits of using agar compared to the lc method for strain isolation?
thanks,
masterplain88

lc doesn't do strain isolation




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