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Doctor Mario
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Registered: 08/07/20
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Re: Trying isolation with agar and petri dishes - thoughts? [Re: WunFunGai]
#26897512 - 08/24/20 05:09 PM (3 years, 5 months ago) |
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Lookin good!
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WunFunGai
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Re: Trying isolation with agar and petri dishes - thoughts? [Re: WunFunGai]
#26900364 - 08/26/20 11:38 AM (3 years, 5 months ago) |
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Update: Jars are moving along, but looks like I might have gotten a hitchhiker on one of my wedges - had a little struggle with that wedge and this only highlights how important it is to be careful and methodical in process.
Not sure if it's a leapoff from jiggling around with the knife, or something else.
Anyone know if strong myc can overcome that little piece or if it'll be a "toss"?
 

Wedges definitely leaping off and grabbing onto grains though, and plates seem to be doing well, but only one looks pretty solid without fuzzies.
WFG
-------------------- The Official TNF Reccomended Teks & Methods Bod's Simplified Cultivation Methods <-- Awesome!!! D3monics Perfect Transfers and Agar Tek No matter where you go, there you are... - Buckaroo Banzai You gotta be here, if you're not all there...
Edited by WunFunGai (09/07/20 10:03 PM)
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Martinsapin
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Re: Trying isolation with agar and petri dishes - thoughts? [Re: WunFunGai]
#26900423 - 08/26/20 12:16 PM (3 years, 5 months ago) |
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Cool project !
IMO that wedge got something more that only the myc. And yes its gonna affect its viability. Id toss it.
I hope that you shake you jars right after Agar2grain.
Im definitely following this thread.
-------------------- looking for a sclerotia producer print
Edited by Martinsapin (08/26/20 12:17 PM)
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WunFunGai
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Re: Trying isolation with agar and petri dishes - thoughts? [Re: Martinsapin]
#26900430 - 08/26/20 12:22 PM (3 years, 5 months ago) |
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Quote:
Martinsapin said: Cool project !
IMO that wedge got something more that only the myc. And yes its gonna affect its viability. Id toss it.
I hope that you shake you jars right after Agar2grain.
Im definitely following this thread.
Thanks for your reply. I was afraid of that 
No, I did not shake after tossing wedges in. I read a lot of the teks but didn't find anything definitive so left it as is.
In the past I've tried doing all sorts of jar tilting gyrations to put one nugget in north, south, east, west corners of jar, slightly buried and against the glass, but it felt like doing one of those damn brain puzzles. And figured I was not only overthinking it, but increasing chances of contams.
Just to throw it out there: Does everyone shake up the jar after tossing in wedges?
Personally I'd like to see progress and prefer to have some piece of agar against the glass to watch for strands and growth.
Appreciate the comments! Will try and throw up some agar porn later today.
WFG
-------------------- The Official TNF Reccomended Teks & Methods Bod's Simplified Cultivation Methods <-- Awesome!!! D3monics Perfect Transfers and Agar Tek No matter where you go, there you are... - Buckaroo Banzai You gotta be here, if you're not all there...
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Martinsapin
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Re: Trying isolation with agar and petri dishes - thoughts? [Re: WunFunGai]
#26900699 - 08/26/20 02:23 PM (3 years, 5 months ago) |
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yeah man no need to dispatch it north south east west.
the standard procedure is to drop it and shake the jar so the myc is in contact with the grains and so it wont dry alone on top of the grains. It will speed up colonisation.
shake it !
-------------------- looking for a sclerotia producer print
Edited by Martinsapin (08/27/20 08:51 AM)
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monkey_accident
fail now learn later


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Re: Trying isolation with agar and petri dishes - thoughts? [Re: Martinsapin]
#26900768 - 08/26/20 02:52 PM (3 years, 5 months ago) |
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Really liking this thread. Keep er' updated.
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JJFrozen
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Re: Trying isolation with agar and petri dishes - thoughts? [Re: monkey_accident]
#26900915 - 08/26/20 04:19 PM (3 years, 5 months ago) |
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The grey is cob I think. Nasty bugger. Iโm starting agar soon, I think this thread just made me decide to use food coloring after all. Really looks cool and makes the samples easier to see.
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WunFunGai
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Re: Trying isolation with agar and petri dishes - thoughts? [Re: Martinsapin]
#26900994 - 08/26/20 05:07 PM (3 years, 5 months ago) |
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Quote:
Martinsapin said: yeah man no need to dispatch it north south east west.
the standard procedure is to drop it and shake the jar so the myc is in contact with the grains and it will dry alone on top of the grains. It will speed up colonisation.
shake it !
Copy that. Gonna shake it like a polaroid pictcha! lol...Sometimes when I do something new I'm either end of the spectrum, too conservative or too nuts..Thanks for confirmation.
Agar was kinda scary, but the results and being able to see stuff clearly (even with condensation) is much easier than my initial 4oz PastyPlates. I feel all scientific and shit. Need a lab coat.
I'm tossing 3 of the 2nd level xfer plates, but figured I'd drop some backlit pics of the 3 that I'll keep to try a 3rd xfer.
Thanks again for comments, much appreciated.
WFG 
Update: Agar Pr0n 08/25
 
-------------------- The Official TNF Reccomended Teks & Methods Bod's Simplified Cultivation Methods <-- Awesome!!! D3monics Perfect Transfers and Agar Tek No matter where you go, there you are... - Buckaroo Banzai You gotta be here, if you're not all there...
Edited by WunFunGai (09/07/20 10:04 PM)
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Doctor Mario
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Registered: 08/07/20
Posts: 1,894
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Re: Trying isolation with agar and petri dishes - thoughts? [Re: WunFunGai]
#26901078 - 08/26/20 05:58 PM (3 years, 5 months ago) |
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Quote:
WunFunGai said:
Anyone know if strong myc can overcome that little piece or if it'll be a "toss"?
Normally you can still get fruits if theres some bacteria but that usually only applies if the bacteria doesn't show up until around the pinning stage. Being that the myc hasn't even began to colonize grain I would suggest to not use that jar. The myc and bacteria are going to be competing for the nutrients in the grain from here on out. That could compromise your whole grow. Its not worth it to try to save it and get a few fruits when you could easily make another jar and have many fruits.
That's too bad that that wedge went contam. Looking like good myc. Its hard to tell from the pics of the transfers that you're keeping but is that bacteria on 2B1 and 2B2? See how it looks blurry like there's a film covering the bottom part of 2B1 and the top left part of 2B2?
The Mycelium doesn't appear to be growing in those sections either which could indicate that it is in fact bacterial and the contam is winning the fight. I could very well be wrong. I've never done agar or cultivated for that matter but I'd hate to be right and not say anything and have you waste time and effort by using them.
You can post those photo's here and here. Someone with a better eye and experience greater than 0 will probably chime in and let you know for certain.
If they are bacterial, it looks like you can still save them by making another transfer from a sector that the bacteria hasn't gotten to yet.
Also, was that sediment in them from the start or is that something new?
Edited by Doctor Mario (08/26/20 06:16 PM)
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WunFunGai
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Re: Trying isolation with agar and petri dishes - thoughts? [Re: Doctor Mario]
#26913037 - 09/02/20 12:57 AM (3 years, 4 months ago) |
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Quote:
Doctor Mario said: Also, was that sediment in them from the start or is that something new?
Thanks for your reply. All input is useful.
Nah, the sediment is normal when I do PHA (Potato Honey Agar), I use Idahoan Potato Flakes, Organic Honey, and agar-agar, so there's always a bit there but not an issue.
I'm looking into getting some LME in the future, but so far I've used this basic PHA for a bit and it's worked well...If it ain't broke, don't fix it. 
Been busy but some quick update photos below...things moving along. Pics below are of each jar on a few days ago:
   
Group photo

Will try and post more later.
Enjoy.
WFG
-------------------- The Official TNF Reccomended Teks & Methods Bod's Simplified Cultivation Methods <-- Awesome!!! D3monics Perfect Transfers and Agar Tek No matter where you go, there you are... - Buckaroo Banzai You gotta be here, if you're not all there...
Edited by WunFunGai (09/07/20 10:04 PM)
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Martinsapin
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Registered: 09/12/16
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Re: Trying isolation with agar and petri dishes - thoughts? [Re: WunFunGai]
#26913250 - 09/02/20 06:55 AM (3 years, 4 months ago) |
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Im gonna follow this thread until I see fruits
-------------------- looking for a sclerotia producer print
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Doctor Mario
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Registered: 08/07/20
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Re: Trying isolation with agar and petri dishes - thoughts? [Re: Martinsapin]
#26913306 - 09/02/20 07:41 AM (3 years, 4 months ago) |
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WunFunGai
Wanderer

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Re: Trying isolation with agar and petri dishes - thoughts? [Re: Doctor Mario]
#26914086 - 09/02/20 04:43 PM (3 years, 4 months ago) |
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Update: After jars were shook based on advice, here's how they looked a few more days after.
   
Then it was time for something new and completely different...a 3rd Xfer experimenting with something introduced in D3monic's Perfect Transfers and Agar:
 
May start another side project but trying to juggle efficiently.
Thanks for the comments and encouragement. Much appreciated.
Enjoy.
WFG
-------------------- The Official TNF Reccomended Teks & Methods Bod's Simplified Cultivation Methods <-- Awesome!!! D3monics Perfect Transfers and Agar Tek No matter where you go, there you are... - Buckaroo Banzai You gotta be here, if you're not all there...
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Doctor Mario
๐๐๐๐๐



Registered: 08/07/20
Posts: 1,894
Loc: ๐บ๐ธ
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Re: Trying isolation with agar and petri dishes - thoughts? [Re: WunFunGai]
#26914095 - 09/02/20 04:50 PM (3 years, 4 months ago) |
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What are you using for the hole punch?
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WunFunGai
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Re: Trying isolation with agar and petri dishes - thoughts? [Re: Doctor Mario]
#26914922 - 09/03/20 04:16 AM (3 years, 4 months ago) |
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Quote:
Doctor Mario said: What are you using for the hole punch?
Found a 2-pack of steel straws with bendy silicone on end for about $4 that was about the right size made of 18/8 food grade steel.
Pulled the bendy off as a backup to the other straw which I'll use for drinking.
Heated it up with a torch, stabbed in agar a couple times to cool, then cut a puck and used my blade to stab the side of the puck, then flip it over in center of a clean agar plate.
Will try and throw some pics up again later.
WFG
-------------------- The Official TNF Reccomended Teks & Methods Bod's Simplified Cultivation Methods <-- Awesome!!! D3monics Perfect Transfers and Agar Tek No matter where you go, there you are... - Buckaroo Banzai You gotta be here, if you're not all there...
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Doctor Mario
๐๐๐๐๐



Registered: 08/07/20
Posts: 1,894
Loc: ๐บ๐ธ
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Re: Trying isolation with agar and petri dishes - thoughts? [Re: WunFunGai]
#26915046 - 09/03/20 07:07 AM (3 years, 4 months ago) |
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Quote:
WunFunGai said:
Quote:
Doctor Mario said: What are you using for the hole punch?
Found a 2-pack of steel straws with bendy silicone on end for about $4 that was about the right size made of 18/8 food grade steel.
Pulled the bendy off as a backup to the other straw which I'll use for drinking.
Heated it up with a torch, stabbed in agar a couple times to cool, then cut a puck and used my blade to stab the side of the puck, then flip it over in center of a clean agar plate.
Will try and throw some pics up again later.
WFG 
I like it. I might just go to the store and see if I can find something similar.
On a side note, when I was getting my supplies I didn't feel like driving 25 minutes to go to wal mart for an xacto-knife. Walgreens is a few blocks from my house so I went there and looked for a scalpel.
I didnt see one anywhere so I asked the girl at the pharmacy section. She looked super weirded out and whispered to the pharmacist, "He wants a scalpel". With wide eyes the pharmacist looked at me and said, "We don't have any!".
I admit it probably looked suspect AF since I was holding a box of face masks, a box of gloves and a bottle of distilled water. I try to avoid going in there now lol.
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Roger Clemency
Smile


Registered: 03/23/20
Posts: 2,005
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Re: Trying isolation with agar and petri dishes - thoughts? [Re: Doctor Mario]
#26915143 - 09/03/20 08:12 AM (3 years, 4 months ago) |
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Lol pharmacy people are nuts sometimes. I had to buy some syringes once while driving through the country. I think I was in Iowa and stopped at a pharmacy, the dude wouldnโt sell me syringes which was crazy to me cuz Iโve never had trouble getting them in states I lived in. He got pissy with me so I left, went to another pharmacy like 10 minutes away and the guy behind the counter was on the phone saying โyeah I think heโs coming in nowโ I was like Iโm getting outta here before the fuzz show up haha. Bastids.
Anyway...nice looking plates most of them. I donโt like including the very edges of agar plates when I go to grain, the piece you had with the gray mold may have gotten contaminated during inoc but itโs also possible a mold spore had gotten in to the edge of the plate soon before you did your inoculations. The edges are the most vulnerable usually.
I wouldnโt worry too much about sectoring or anything right now, just a few transfers away from spores you still have 3,432 strains there on each plate. Or some number anyway ; )
Once I get fully clean plates (no obvious contams or bald spots of myc or odd growth patterns) Iโll transfer once more and then use the plates for Blenderless LI - https://www.shroomery.org/forums/showflat.php/Number/22833314 It works well and gives you about 10 day colonization of qt jars.
-------------------- Sour grapes, sweet revenge Heaven starts right where hell ends
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WunFunGai
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Re: Trying isolation with agar and petri dishes - thoughts? [Re: Doctor Mario]
#26916135 - 09/03/20 06:09 PM (3 years, 4 months ago) |
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Quote:
Doctor Mario said:
Quote:
WunFunGai said:
Quote:
Doctor Mario said: What are you using for the hole punch?
Found a 2-pack of steel straws with bendy silicone on end for about $4 that was about the right size made of 18/8 food grade steel.
Pulled the bendy off as a backup to the other straw which I'll use for drinking.
Heated it up with a torch, stabbed in agar a couple times to cool, then cut a puck and used my blade to stab the side of the puck, then flip it over in center of a clean agar plate.
Will try and throw some pics up again later.
WFG 
I like it. I might just go to the store and see if I can find something similar.
On a side note, when I was getting my supplies I didn't feel like driving 25 minutes to go to wal mart for an xacto-knife. Walgreens is a few blocks from my house so I went there and looked for a scalpel.
I didnt see one anywhere so I asked the girl at the pharmacy section. She looked super weirded out and whispered to the pharmacist, "He wants a scalpel". With wide eyes the pharmacist looked at me and said, "We don't have any!".
I admit it probably looked suspect AF since I was holding a box of face masks, a box of gloves and a bottle of distilled water. I try to avoid going in there now lol.
Hey, these work great:
63 Pack - 3 Exacto Knives and 60 Blades (6pks of 10) - https://www.amazon.com/gp/product/B07VZTMKXK/ref=ppx_yo_dt_b_asin_title_o06_s00?ie=UTF8&psc=1
Innoculating Loops (50pcs) and holder- https://www.amazon.com/gp/product/B074M9QQ64/ref=ppx_yo_dt_b_asin_title_o06_s00?ie=UTF8&psc=1
Both under $10 each...good additions to toolkit. Just wish the handle/rod on innoc loop was a bit shorter.
Hope this helps.
WFG
-------------------- The Official TNF Reccomended Teks & Methods Bod's Simplified Cultivation Methods <-- Awesome!!! D3monics Perfect Transfers and Agar Tek No matter where you go, there you are... - Buckaroo Banzai You gotta be here, if you're not all there...
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Doctor Mario
๐๐๐๐๐



Registered: 08/07/20
Posts: 1,894
Loc: ๐บ๐ธ
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Re: Trying isolation with agar and petri dishes - thoughts? [Re: WunFunGai]
#26916320 - 09/03/20 08:12 PM (3 years, 4 months ago) |
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Thank you. Im definitely gonna get the loops. I made one out of aluminum wire and I dont care for it much.
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lookintolearn
Stranger becoming Strangest



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Re: Trying isolation with agar and petri dishes - thoughts? [Re: Doctor Mario]
#26916341 - 09/03/20 08:26 PM (3 years, 4 months ago) |
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I started using D3's leather hole punch idea for transfers and i'm never going back. It makes it so much easier to get beautiful agar and it's so much easier to get the wedge onto the transfer plate, with normal wedges I always had trouble getting it off the scalpel but with these you can just gently lift it from the side, flip it, slides right off.
-------------------- Don't be afraid of feeling the feelings Lookin to LAGM 2021    Looking to start growing? Read through Bod's Introduction to Everything Looking to start agar? Start with Alien's Holy Grail Looking to perfect your transfers? Start with D3monic's Perfect Transfers Looking for easiest prep to Coir ever? Eat's UNBUCKET Tek Looking to start LC? Try LI first! Munch's super easy Blenderless LI
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