Agar Poke-a-Plates

Sideshow · 08-25-09, 17:06

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.
http://forums.mycotopia.net/agar-str...zai-style.html (Agar for the masses - Banzai Style!)

http://forums.mycotopia.net/air-circ...-airports.html (BB does airports)

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:

This is what the finished plate looks like:

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)

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

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

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

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

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!

Sideshow · 08-25-09, 17:09

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

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

Elf Salvation · 08-25-09, 18:11

trying this one out myself

Foster · 08-25-09, 20:00

Nice work man.

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.

roscoe · 08-25-09, 20:18

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

http://forums.mycotopia.net/attachme...es-syringe.jpg

love this pic!

Ben Dover · 08-25-09, 20:36

Quote:

Originally Posted by roscoe

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

Sideshow · 08-25-09, 22:19

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.

roscoe · 08-26-09, 04:30

Quote:

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.

Sideshow · 08-29-09, 00:14

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

A control (one of the 9 done before)

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

bozefonce · 08-29-09, 00:25

Quote:

Originally Posted by Sideshow

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!

masterplain88 · 08-29-09, 00:28

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

Sideshow · 08-29-09, 00:32

Quote:

Originally Posted by masterplain88

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://forums.mycotopia.net/cloning/...ek-merged.html (Buckaroo's Cloning Tek...{merged})

masterplain88 · 08-29-09, 00:37

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

Sideshow · 08-29-09, 00:42

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

Leftover Salmon · 08-29-09, 01:06

Quote:

Originally Posted by Sideshow

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.

weeeeeee · 08-29-09, 01:35

Quote:

Originally Posted by roscoe

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

http://forums.mycotopia.net/attachme...es-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

Sideshow · 08-29-09, 09:06

Quote:

Originally Posted by Leftover Salmon

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.

BuckarooBanzai · 08-29-09, 11:47

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.

Sideshow · 08-29-09, 12:33

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.

Hippie3 · 08-29-09, 14:32

Quote:

Originally Posted by masterplain88

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

Hippie3 · 08-29-09, 14:33

Quote:

Originally Posted by Hippie3

lc doesn't do strain isolation

pure genius, sideshow.

masterplain88 · 08-29-09, 14:35

yea thats what i thought but then i saw the lc part. thanks for clearing that up hippie3. 08-29-09 00:14SideshowUpdate
3 days later, we have some myc growth from 2 lc's:

Attachment 139604

Attachment 139605

A control (one of the 9 done before)

Attachment 139606

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

Attachment 139607

Sideshow · 08-29-09, 14:38

That means a lot to me Hippie, thank you.

masterplain88 · 08-29-09, 14:40

sorry do be a bother but what is the ratio and mix procedures for your mrya malt agar.? thanks,
im thinking of just buying pr-made agar.
masterplain88

Quote:

Originally Posted by Sideshow

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://forums.mycotopia.net/cloning/...ek-merged.html (Buckaroo's Cloning Tek...{merged})

Sideshow · 08-29-09, 15:03

Quote:

Originally Posted by masterplain88

sorry do be a bother but what is the ratio and mix procedures for your mrya malt agar.? thanks,
im thinking of just buying pr-made agar.
masterplain88

I would reccommend premixed powder agar if it is your first time. Sporeworks brand PDA is great, for one.

MRYA Recipe: (1/2 pint)
250ml Water
1.5g BRF
3g Malt Extract
.25g Nutritional Yeast
3g Agar

I used to premix the ingredients in boiling water and then pc, but that is a PITA. Nowadays, I just dump everything into a pint mason, shake vigorously for a minute, and pc 30 minutes. Lastly, I also make sure that the agar has not separated (by lightly swirling the jar) as it hardens.

masterplain88 · 08-29-09, 15:29

thanks bro good write up you got here for some one like me looking to get into isolation work. your a true asset to the topia community. good work man.
peace.
masterplain88

Leftover Salmon · 08-29-09, 15:38

One more newbish question from Salmon,,, Could swim use an isolated chunk of myc growth from agar to make one stellar LC? Or would this be a waste of time? Also could swim drop pieces of agar with myc directly into a grain jar or bag? Thanks you for the knowledge!

Sideshow · 08-29-09, 16:14

Quote:

Originally Posted by Leftover Salmon

Could swim use an isolated chunk of myc growth from agar to make one stellar LC?

That's exactly what I do

Quote:

Originally Posted by Leftover Salmon

could swim drop pieces of agar with myc directly into a grain jar or bag?

Yessir

BuckarooBanzai · 08-29-09, 17:45

When agar is well colonized, it is easy to transfer myc with a syringe of sterile water.

Just open the plate enough to stick the needle in and then squirt in 2-3mL of water. The myc is very easy to scrape off the agar with the sterile needle and then you can suck the myc water back up into the syringe.

I really prefer to do myc transfers as LCs or liquid slurry in syringes. Syringes cut contam rates compared to open jars and plates, even with a flow hood.

And you can do syringe work very successfully without a flow hood.

Leftover Salmon · 08-29-09, 22:26

Awesome! Thanks for the tips folk! I, Swim, and FOAF all love this place & the people within.

I am excited to try this agar biz...

bugs · 08-29-09, 23:34

Quote:

Originally Posted by BuckarooBanzai

When agar is well colonized, it is easy to transfer myc with a syringe of sterile water.

Just open the plate enough to stick the needle in and then squirt in 2-3mL of water. The myc is very easy to scrape off the agar with the sterile needle and then you can suck the myc water back up into the syringe.

Semms that would work fine in the cuppy things, too. Poke a needle throuhg the lid, drip and sip.

By the way, frickin' ingenious tek. Gotta love that high-grade ghetto. Perfect match for the shmuvbox.

masterplain88 · 09-08-09, 16:32

procedure on the tek would work with bub tweeks like

Quote:

Originally Posted by bugs

Semms that would work fine in the cuppy things, too. Poke a needle throuhg the lid, drip and sip.

By the way, frickin' ingenious tek. Gotta love that high-grade ghetto. Perfect match for the shmuvbox.

waterchilde · 11-06-09, 08:28

Hey, found a source for needles that has 14 ga on up and up to 3' needles. Howard electronic instruments is online and I have their ph # , 1-800-394-1984 . now, can you tell me if the nylon vet syringes are autoclavable or is there a way that I can re-sterilize these new syringes over again for multiple useage. looking to learn thanx...

waterchilde · 11-06-09, 08:32

found it! fo all needle supplies, check this out. http://www.howardelectronics.com/jensen/needles.html

Indonesia · 11-06-09, 11:19

Nice work sideshow. my foaf has done this tek before with petris..good sound agar technique for a glovebox.

copelandiaKidd · 11-06-09, 12:16

this is awesome!!!! I have been looking for an alternative to half pints for a while now!!!!!

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08-25-09, 17:06	#1 (permalink)
Sideshow Freakishly Large Join Date: Sep 2007 Posts: 353	Agar Poke-a-Plates 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. http://forums.mycotopia.net/agar-str...zai-style.html (Agar for the masses - Banzai Style!) http://forums.mycotopia.net/air-circ...-airports.html (BB does airports) 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: This is what the finished plate looks like: 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) I take my prepared agar from storage and heat it in a water bath. And suck it into a big syringe. 10ml syringes can get you 2 plates if you are stingy. Then I wipe the lid with alcohol and inject the agar. 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 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 __________________ Thank you, Mr. Mason

08-25-09, 17:09	#2 (permalink)
Sideshow Freakishly Large Join Date: Sep 2007 Posts: 353	Oh and you can get the cups here: http://www.wesellcoffee.com/page29.html The ones I use are 2 oz, but some may like the 3.25oz better. __________________ Thank you, Mr. Mason

08-25-09, 18:11	#3 (permalink)
Elf Salvation VIP Member Join Date: Nov 2005 Posts: 621	trying this one out myself __________________ Hey...... can it stop

08-25-09, 20:18	#5 (permalink)
roscoe Smother? Join Date: Oct 2008 Posts: 233	i have no expirence with agar yet. but i read a clean work station is paramount. maybe not? http://forums.mycotopia.net/attachme...es-syringe.jpg love this pic! __________________ Don't believe everything I think.

08-25-09, 20:00	#4 (permalink)
Foster intotheoutside Join Date: Jun 2006 Posts: 1,830	Nice work man. 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.

08-25-09, 22:19	#7 (permalink)
Sideshow Freakishly Large Join Date: Sep 2007 Posts: 353	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. __________________ Thank you, Mr. Mason

08-29-09, 00:14	#9 (permalink)
Sideshow Freakishly Large Join Date: Sep 2007 Posts: 353	Update 3 days later, we have some myc growth from 2 lc's: A control (one of the 9 done before) And one that isn't looking well (the only one that has gone south) __________________ Thank you, Mr. Mason

08-29-09, 00:28	#11 (permalink)
masterplain88 Mycotopiate Join Date: Jan 2009 Posts: 683	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

08-29-09, 00:37	#13 (permalink)
masterplain88 Mycotopiate Join Date: Jan 2009 Posts: 683	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

08-29-09, 00:42	#14 (permalink)
Sideshow Freakishly Large Join Date: Sep 2007 Posts: 353	2 advantages: Agar is the only way to clean up a culture and Agar allows more reliable substrain isolation than other methods, IMO __________________ Thank you, Mr. Mason

08-29-09, 11:47	#18 (permalink)
BuckarooBanzai Prone to ranting... Join Date: Oct 2005 Posts: 6,050	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. __________________ Banzai Institute for Higher Education (a collection of growing Teks & threads)

08-29-09, 12:33	#19 (permalink)
Sideshow Freakishly Large Join Date: Sep 2007 Posts: 353	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. __________________ Thank you, Mr. Mason

08-29-09, 14:35	#22 (permalink)
masterplain88 Mycotopiate Join Date: Jan 2009 Posts: 683	yea thats what i thought but then i saw the lc part. thanks for clearing that up hippie3. 08-29-09 00:14SideshowUpdate 3 days later, we have some myc growth from 2 lc's: Attachment 139604 Attachment 139605 A control (one of the 9 done before) Attachment 139606 And one that isn't looking well (the only one that has gone south) Attachment 139607

08-29-09, 14:38	#23 (permalink)
Sideshow Freakishly Large Join Date: Sep 2007 Posts: 353	That means a lot to me Hippie, thank you. __________________ Thank you, Mr. Mason

08-29-09, 15:29	#26 (permalink)
masterplain88 Mycotopiate Join Date: Jan 2009 Posts: 683	thanks bro good write up you got here for some one like me looking to get into isolation work. your a true asset to the topia community. good work man. peace. masterplain88

08-29-09, 15:38	#27 (permalink)
Leftover Salmon Can'tGetRight Join Date: Aug 2009 Posts: 224	One more newbish question from Salmon,,, Could swim use an isolated chunk of myc growth from agar to make one stellar LC? Or would this be a waste of time? Also could swim drop pieces of agar with myc directly into a grain jar or bag? Thanks you for the knowledge!

08-29-09, 17:45	#29 (permalink)
BuckarooBanzai Prone to ranting... Join Date: Oct 2005 Posts: 6,050	When agar is well colonized, it is easy to transfer myc with a syringe of sterile water. Just open the plate enough to stick the needle in and then squirt in 2-3mL of water. The myc is very easy to scrape off the agar with the sterile needle and then you can suck the myc water back up into the syringe. I really prefer to do myc transfers as LCs or liquid slurry in syringes. Syringes cut contam rates compared to open jars and plates, even with a flow hood. And you can do syringe work very successfully without a flow hood. __________________ Banzai Institute for Higher Education (a collection of growing Teks & threads)

08-29-09, 22:26	#30 (permalink)
Leftover Salmon Can'tGetRight Join Date: Aug 2009 Posts: 224	Awesome! Thanks for the tips folk! I, Swim, and FOAF all love this place & the people within. I am excited to try this agar biz...

11-06-09, 08:28	#33 (permalink)
waterchilde Mycophiliac Join Date: Nov 2009 Posts: 9	luer lock needles Hey, found a source for needles that has 14 ga on up and up to 3' needles. Howard electronic instruments is online and I have their ph # , 1-800-394-1984 . now, can you tell me if the nylon vet syringes are autoclavable or is there a way that I can re-sterilize these new syringes over again for multiple useage. looking to learn thanx...

11-06-09, 08:32	#34 (permalink)
waterchilde Mycophiliac Join Date: Nov 2009 Posts: 9	here it is found it! fo all needle supplies, check this out. http://www.howardelectronics.com/jensen/needles.html

11-06-09, 11:19	#35 (permalink)
Indonesia Mycotopiate Join Date: Nov 2007 Posts: 217	Nice work sideshow. my foaf has done this tek before with petris..good sound agar technique for a glovebox.

11-06-09, 12:16	#36 (permalink)
copelandiaKidd Void Of Boundaries Join Date: Jul 2009 Posts: 578	this is awesome!!!! I have been looking for an alternative to half pints for a while now!!!!! __________________ "LEARN THE RULES SO YOU CAN BREAK THEM PROPERLY" -The Dali lama "i don't care if it eats my cat, as long as the damn thing fruits"~ prism

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