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OfflineWunFunGai
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Registered: 05/03/16
Posts: 165
Loc: Skull Island
Last seen: 15 days, 20 hours
Isolating with agar and petri dishes - A walk down isolation road * 1
    #26889280 - 08/19/20 11:19 PM (3 years, 5 months ago)

Greetings! It's been awhile since my last post, but got the itch after a break to revive the hobby and try something new since I've got a bit of time on my hands.

First pass was over two decades ago stumbling upon the original mmgg.html doc and getting my first spores from PF back in the day.

Flash forward to 2016 and revisited the tek and improvements and overwhelming amount of posts with info. Started from old roots; went brf and shorties in a SGFC, moved on to shoe-boxes, then unmodified tubs based on the postings of The Bod...got a cycle going and was having fun just repeating the process based on a clone of a good fruit and using pasty plates.

Had to move and had some unfortunate contam issues and put things on the side until the present and decided to dive in again and learn how to do real agar plates and pour agar and see if any old plates had life left and take my first shot at isolating starting by going spores to agar.

And here's where I'm at...followed info I found and got some 500ml media bottles with GL45 lids and whipped up some PDA using agar-agar, potato flakes, and honey w/ food coloring. PC'd the mix for 30 minutes, let it cool, then used short microwave bursts to get it liquid again before pouring out 20 plates using the guides that Bod had organized.

NOTE: The spores are Golden Teacher from fruit that was spawned from a good clone from long ago.

Aside from some condensation issues (I know better now), plates went well and on 08/5 I swabbed two plates, and used two others to see if old agar in fridge was viable...old agar was a bust, but on 08/13 the spore plates looked like this:



On 08/14 I culled 3 samples from each plate and put them to 3 new plates:



NOTE: The C Plates look that way since I was lazy and dropped the culled sample on top of my old agar cultures that were doing nothing.

08/16 here's how they look:



08/17 and lots of action:
Source plate 01 starting to look weird (toss?)


1A - looking good but is that unwanted visitor at 4 o clock? Started small but it's rapidly outpacing on 08/19


2B - Looking good is that 3 o clock section a different sector?


3C - Looking OK too


PLATE 02 - looking kinda beat up now (toss?)


2A - Looks OK, but kinda meh for speed


2B - Looks pretty good - that 3 o clock section looks interesting


2C - Looks OK, but that whole lower half seems a bit sketchy.


I'm still a noob, I've read a lot of different info, and I'm a bit overwhelmed by some of the conflicting info and just looking for comments, suggestions, encouragement, or even ridicule just to get perspective.

I'm just hoping to make fewer mistakes on the way to learning proper isolation techniques. I'll probably make some anyway, but the goal is to make fewer...

* I've burned 8 plates have twelve poured left in storage.
* I have two sleeves of petris 20 plates like shown, and 20 Y plates same size that are unused.

I'm thinking of taking at 1-2 samples of best growth from each plate and grow those out while starting another bottle of agar and dishes.

If the new plates don't look isolated enough, I may put to new plates and experiment with the leftovers to experiment and see what comes out.

If anyone has followed the wall of text this far, I've tried to provide a decent description of what I've done and where I'm at and would greatly appreciate any comments regarding next step, or if you have a good eye for spotting sectors to point out what a more experienced person would do.

Please be patient. This is my first post outside of The Noob Forum and I'm a bit intimidated by all the more experienced posters.

Any comments would be appreciated, and in turn I'll post updates as I walk down isolation road...

Thanks in advance for any thoughts, info, or links.

WFG :smile:


--------------------
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 (09/10/20 02:23 AM)


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OfflineWunFunGai
Wanderer

Registered: 05/03/16
Posts: 165
Loc: Skull Island
Last seen: 15 days, 20 hours
Re: First timer trying isolation with agar and petri dishes - thoughts? [Re: WunFunGai]
    #26890109 - 08/20/20 01:34 PM (3 years, 5 months ago)

Just figured I'd provide a quick update. No responses, but maybe this info will be useful for others and maybe spare them any mistakes I'm making along the way.

08/20 - Main Plates look pretty burned out - is it normal for myc to eat the food coloring away like that?



Plate 1 Transfers: Noted that funky spot going nuts on 1A - gray stuff ain't good right?


Plate 2 Transfers: Got a funky spot on one, but I think it's just myc eating around the old agar that didn't do anything.


Pulled 6 plates from fridge and "uncondensing" them using the hot cup trick.

Wondering if I should take one good sample from each plate or focus on one and put the others in cold storage to slow further growth.

My plan was to try and get at least 4 plates that looked really good, then spawn each to 1 quart of grain and shoebox it to see how they come out.

If there's exceptional results from any of the four, make more plates from that, and try a larger run.

I'll just continue talking to myself here since putting it in private logs probably won't help (or amuse) anyone.

Comments or input much appreciated, just trying to make less mistakes and optimize my process where possible.

Have a good one!

WFG :smile:


--------------------
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 10:05 PM)


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OfflineWunFunGai
Wanderer

Registered: 05/03/16
Posts: 165
Loc: Skull Island
Last seen: 15 days, 20 hours
Re: First timer trying isolation with agar and petri dishes - thoughts? [Re: WunFunGai]
    #26892160 - 08/21/20 05:09 PM (3 years, 5 months ago)

Update 08/21 - No responses so moving along...

The master plates don't look so hot and will toss those, and of the 3 remaining plates, 1A has something in the that I don't think is myc. 1C/2C look ok, but are growing around the old agar chunks that didn't produce - the fuzzy stuff in those dead sections worry me a bit, but put em in a ziploc and put them in cold storage for now.

Conclusion: I guess myc does eat food coloring after all..



Decided to take the 3 best looking plates and cull two samples from each and put them to plates.

Here's the 3 that were culled.



Figured wtf, I got some time on my hands so I started a grain soak, fill 4x1qt jars and PC them, then noc em up with small nuggets of agar from my 3 nice plates I just took samples from to isolate with.

Just for shits and giggles while I wait for the 6 new plates to grow out.

Figger I'll spawn them to 4 shoebox runs to keep them separate and observe any differences.

I'm pretty much going with what I've read from various teks but feeling nervous about what I may be missing or could be doing better.

Comments and suggestions appreciated, even if it's just to confirm my process is OK and things aren't looking bad yet.

Have a good one...

WFG :smile:


--------------------
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 10:05 PM)


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OfflineOne of Us
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Registered: 03/12/12
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Re: First timer trying isolation with agar and petri dishes - thoughts? [Re: WunFunGai]
    #26892314 - 08/21/20 07:07 PM (3 years, 5 months ago)

Everything looks great! Good job!


--------------------
LAGM 2021
LAGM 2020
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Offlineredbee
prole spore threat
Female


Registered: 03/30/20
Posts: 175
Last seen: 1 year, 8 months
Re: First timer trying isolation with agar and petri dishes - thoughts? [Re: WunFunGai]
    #26892337 - 08/21/20 07:23 PM (3 years, 5 months ago)

I can't offer you any advice as I'm new to agar, but I just wanted to say thanks for the thorough write-up, it's really helpful to see your process!

Good luck with those grain jars, and I'm staying tuned...
:popcorn:


--------------------
:sporedrop:  redbee's LAGM 2021  :sporedrop:


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OfflineWunFunGai
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Registered: 05/03/16
Posts: 165
Loc: Skull Island
Last seen: 15 days, 20 hours
Re: First timer trying isolation with agar and petri dishes - thoughts? [Re: One of Us]
    #26892733 - 08/22/20 12:53 AM (3 years, 5 months ago)

Quote:

One of Us said:
Everything looks great! Good job!




Hey, thanks! I hit up some of your LAGM posts too, and I'm glad to see I'm not alone with condensation on plates! I've been away for awhile.

Local environment is hot/humid so stuff happens fast once it starts.

My hypothesis is that although myc is pretty strong and resilient when established. It's kryptonite is warmer temps, which is the supersauce environment for Mr. Grey and w/humid conditions he wins in the end...major clash of titans action at the demarcation on that plate eh? (Sorry, said with a science background it might sound less interesting than it could be...right?)...Two Strains, One Plate, There Can Be Only One.....TOSS...LOL

Thanks for your comments. Really appreciated!

WFG :smile:


--------------------
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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OfflineWunFunGai
Wanderer

Registered: 05/03/16
Posts: 165
Loc: Skull Island
Last seen: 15 days, 20 hours
Re: First timer trying isolation with agar and petri dishes - thoughts? [Re: redbee]
    #26892736 - 08/22/20 12:57 AM (3 years, 5 months ago)

Hey, thanks!

I hope it's helpful to add to other good info already here to give some perspective.

This is just my interpretation, I hope it adds/helps your process, don't forget, do what works for you! :smile:

Thanks,

WFG :smile:


--------------------
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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OfflineSteelGully
Just Strange
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Registered: 06/02/20
Posts: 64
Loc: Bottom of the world
Last seen: 3 years, 4 months
Re: First timer trying isolation with agar and petri dishes - thoughts? [Re: WunFunGai]
    #26892785 - 08/22/20 02:08 AM (3 years, 5 months ago)

Looks epic! I'm still learning too so this is super valuable follow along with epic pics and great insight on what you are doing. Thanks for sharing sorry I have nothing helpful to add.


--------------------
Loving the shroom journey!
:mushroom2: :nyan: :tardpig: :goat: :sporedrop:
:typonese: :aweyeah:


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OfflineJatzan
Mycophile
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Registered: 07/21/20
Posts: 148
Last seen: 2 years, 4 months
Re: First timer trying isolation with agar and petri dishes - thoughts? [Re: SteelGully]
    #26892800 - 08/22/20 02:23 AM (3 years, 5 months ago)

:leocheers:
Amazing journal, thorough research, meticulous work, good technique, flawless photos, modest character. Good job and Keep up!


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OfflineWunFunGai
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Registered: 05/03/16
Posts: 165
Loc: Skull Island
Last seen: 15 days, 20 hours
Re: Trying isolation with agar and petri dishes - thoughts? [Re: WunFunGai]
    #26893620 - 08/22/20 01:35 PM (3 years, 5 months ago)

Thanks for the kind words everyone. Gonna PC up some grain today...

Here's some backlit shots of the 3 I culled from and tried to adjust to get a good view of the strands.

Musing out loud here, but I just haven't developed a good eye for identifying sectors.

I may try drawing in what I think are sectors later and sharing and see if anyone with a good eye will comment.



Enjoy your day!

WFG :smile:


--------------------
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 10:06 PM)


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InvisibleDoctor Mario
πŸ„πŸ„πŸ„πŸ„πŸ„
Male Unread Journal


Registered: 08/07/20
Posts: 1,894
Loc: πŸ‡ΊπŸ‡Έ
Trusted Cultivator
Re: Trying isolation with agar and petri dishes - thoughts? [Re: WunFunGai]
    #26893655 - 08/22/20 02:11 PM (3 years, 5 months ago)

Thanks for sharing. I'll be starting agar soon and I enjoy following along. Keep em comin.


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OfflineJatzan
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Registered: 07/21/20
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Re: Trying isolation with agar and petri dishes - thoughts? [Re: WunFunGai]
    #26893957 - 08/22/20 05:33 PM (3 years, 5 months ago)

Quote:

WunFunGai said:


Enjoy your day!





This is some serious agar porn! So perfect, so beautiful... daaaamn.


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OfflineYumyumyumyum
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Registered: 12/21/19
Posts: 412
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Re: First timer trying isolation with agar and petri dishes - thoughts? [Re: One of Us]
    #26893996 - 08/22/20 05:53 PM (3 years, 5 months ago)

It is tough sometimes with the replies. For a starter those plates look well made. The samples are nice. Agar has not been too kind to me. Out of the 20 odd plates I have made only 4 have been viable and I never realized you cut out the good stuff and transfer it. The samples do look nice from what I understand about the subject. I make that assumption based on the strand lengths.


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OfflineWunFunGai
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Registered: 05/03/16
Posts: 165
Loc: Skull Island
Last seen: 15 days, 20 hours
Re: Trying isolation with agar and petri dishes - thoughts? [Re: Jatzan]
    #26894652 - 08/23/20 12:44 AM (3 years, 5 months ago)

Quote:

Jatzan said:
This is some serious agar porn! So perfect, so beautiful... daaaamn.




Thanks! And thanks for my first ever rate!! :smile:

I thought you might enjoy those so I figured I'd share to the thread.

Meanwhile back at the ranch...



Bout ready PC up some...grainnnnssss (worst zombie impression ever)

G'night Shroomerites! Till the morrow!

WFG :smile:


--------------------
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 10:06 PM)


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InvisibleDoctor Mario
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Registered: 08/07/20
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Trusted Cultivator
Re: Trying isolation with agar and petri dishes - thoughts? [Re: WunFunGai]
    #26894985 - 08/23/20 08:41 AM (3 years, 5 months ago)

:zombie4: ...grainnnnssss

How do you like those plastic lids? I was gonna get some but the only place I found them wanted almost 13 dollars for 6 lids. Thats more than what I paid for the jars.


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OfflineWunFunGai
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Registered: 05/03/16
Posts: 165
Loc: Skull Island
Last seen: 15 days, 20 hours
Re: Trying isolation with agar and petri dishes - thoughts? [Re: Doctor Mario]
    #26895396 - 08/23/20 01:16 PM (3 years, 5 months ago)

Quote:

Doctor Mario said:
:zombie4: ...grainnnnssss

How do you like those plastic lids? I was gonna get some but the only place I found them wanted almost 13 dollars for 6 lids. Thats more than what I paid for the jars.




I like them a shit ton better than the regular white one's which crack easy and  seem pretty lightweight compared to these. The white one's also seemed to deform/deteriorate faster after multiple uses.

I bought two boxes (6 lids each) for $4.49/bx at Target in January 2020 just as the rona was ramping up. $9 for 12 was a great deal! I only typically run 4 qts at a time in two 20qt unmodded tubs so that's sufficient to have stuff lined up or stored if I wanted.

For reference - UPC: 014400108136 -

A quick online search turns up plenty links, but most are either out of stock or overpriced IMHO. Looks like they may be making them from Unobtanium these days :undecided: Target still lists them for $4.49, but they've shown out of stock every time I've tried since January.

TBH, I'd reluctantly pay $13 for a box if I could find them because the quality is that good...and grey/silver/blue are my favorite colors. LOL

I've also checked Amazon for similar and I almost pulled the trigger on some bright colored ones that had straw holes already in them to eliminate the whole PITA drilling part...the grey lids are not as brittle and much easier to drill without cracking. I look for lids sized for Wide Mouth Mason Jars.

Hope this info is useful.

WFG :smile:


--------------------
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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InvisibleDoctor Mario
πŸ„πŸ„πŸ„πŸ„πŸ„
Male Unread Journal


Registered: 08/07/20
Posts: 1,894
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Re: Trying isolation with agar and petri dishes - thoughts? [Re: WunFunGai]
    #26895600 - 08/23/20 03:20 PM (3 years, 5 months ago)

I'll probably wait until my metal ones start to rust and then buy all plastic. Thank you.


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OfflineWunFunGai
Wanderer

Registered: 05/03/16
Posts: 165
Loc: Skull Island
Last seen: 15 days, 20 hours
Re: Trying isolation with agar and petri dishes - thoughts? [Re: Doctor Mario]
    #26895649 - 08/23/20 04:04 PM (3 years, 5 months ago)

Quote:

Doctor Mario said:
I'll probably wait until my metal ones start to rust and then buy all plastic. Thank you.




Gotcha. If it ain't broke, don't fix it. Not as much of an option in a humid/salt air laden environment - rust is alive and kicking everywhere. So transitioned to plastic pretty quickly.

While I'm posting, I figured I'd throw in a mini-update on the 2nd transfers of the 3 plates I culled:







That #2 plate seems to have performed well across the board so far.

Gonna see about noccing up some jars with some agar from 1st Xfer to see what happens...

Have a good one!

WFG :smile:


--------------------
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 10:07 PM)


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InvisibleDoctor Mario
πŸ„πŸ„πŸ„πŸ„πŸ„
Male Unread Journal


Registered: 08/07/20
Posts: 1,894
Loc: πŸ‡ΊπŸ‡Έ
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Re: Trying isolation with agar and petri dishes - thoughts? [Re: WunFunGai]
    #26895731 - 08/23/20 05:14 PM (3 years, 5 months ago)

Quote:

WunFunGai said:

Gotcha. If it ain't broke, don't fix it. Not as much of an option in a humid/salt air laden environment - rust is alive and kicking everywhere.

That #2 plate seems to have performed well across the board so far.

Gonna see about noccing up some jars with some agar from 1st Xfer to see what happens...




Its very humid where I live too but im far enough from the coast that rust isn't a real issue. I lived close to Salt Lake city for a while where everything is rust.

Plate 2b1 has some real nice sectoring goin on. You gonna transfer it any further? Could even take a transfer from each sector and see how they perform individually.

Im jealous. Cant wait to start agar. Super paranoid about sterile tek and a little nervous buy excited nonetheless.

Keep us updated please. Ive enjoyed following a long this far.


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OfflineWunFunGai
Wanderer

Registered: 05/03/16
Posts: 165
Loc: Skull Island
Last seen: 15 days, 20 hours
Re: Trying isolation with agar and petri dishes - thoughts? [Re: Doctor Mario]
    #26897233 - 08/24/20 02:11 PM (3 years, 5 months ago)

Update: Another step down iso road...PCd 4 jars and noccd up with 1st xfers that looked I thought looked sexay...will try and post updates of 2nd xfers later.



Nocc'd & Loaded...

WFG :smile:


--------------------
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 10:07 PM)


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