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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]
    #26916646 - 09/04/20 12:26 AM (3 years, 5 months ago)

Quote:

Doctor Mario said:
Thank you. Im definitely gonna get the loops. I made one out of aluminum wire and I dont care for it much.




I've used stainless steel paper clips and a needle nose plier to wrap them, but this is much easier.

It's a bit frail for my tastes but it worked good based on what I've posted in my isolation log for doing both a zig zag and rotational swab as noted in the logs.

Hope it works for you as well!

Be safe, be well...

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
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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]
    #26917404 - 09/04/20 12:52 PM (3 years, 5 months ago)

Update: Things moving along on the plates that were xferred, jars looking almost ready too - will try and put up more pics later today or this weekend.

3rd Xfer taken on 08/31 below:


Enjoy.

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 10:03 PM)


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Invisiblefilthyknees
no coincidence
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Registered: 03/08/13
Posts: 6,283
Trusted Cultivator
Re: Trying isolation with agar and petri dishes - thoughts? [Re: WunFunGai]
    #26917409 - 09/04/20 12:54 PM (3 years, 5 months ago)

:popcorn:


--------------------
But if you're in a hurry, and really got to go
If you're in a hurry, might have to find out slow
That it's one thing to try and another to fly
You get there quicker just a step at a time
It's one thing to bark, another to bite
The show ain't over till you pack up at night


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Offlinemushman1017
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Registered: 08/27/19
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Re: Trying isolation with agar and petri dishes - thoughts? [Re: filthyknees]
    #26917434 - 09/04/20 01:14 PM (3 years, 5 months ago)

What agar recipe are you using? Ive been doing a 2% lme and 2% agar and my shit has been really tomentose lately. :frown:


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OfflineMartinsapin
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Registered: 09/12/16
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Re: Trying isolation with agar and petri dishes - thoughts? [Re: mushman1017]
    #26917458 - 09/04/20 01:23 PM (3 years, 5 months ago)

Wow those are nice plates!
Yeah the d3monic way with the punch...idk if it makes a big difference vs working with a scalpel? What you guys think?


--------------------
looking for a sclerotia producer print


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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: mushman1017]
    #26918868 - 09/05/20 08:58 AM (3 years, 5 months ago)

Quote:

mushman1017 said:
What agar recipe are you using? Ive been doing a 2% lme and 2% agar and my shit has been really tomentose lately. :frown:




Nothing fancy, just PHA (Potato Flakes, Honey, Agar) I wish I took notes on this one, as this is the first time using this combo. Previously for my PastyPlates I used Potato Flakes, Karo, Agar.

It kinda went like this...On top of old horror hill in a secret laboratory, Professor Weirdo and Count Kook were in their monstrous glory. 6 drops of the essence of terror, 5 drops of sinister sauce...oh wait. Wrong flashback. LOL

For 500ml I believe I used:
5g Idahoan Potato Flakes
10g Organic Honey
9g agar-agar

5-6 drops of designer food coloring of your choice - I like the color for contrast to spot contams, but I can see using different colors to mark batches of plates too.

I'm doing another flask of agar and will take better notes.

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
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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: Martinsapin]
    #26918928 - 09/05/20 09:38 AM (3 years, 5 months ago)

Quote:

Martinsapin said:
Wow those are nice plates!
Yeah the d3monic way with the punch...idk if it makes a big difference vs working with a scalpel? What you guys think?




For me, I would say yes.

I spotted the link at the end of Doctor Mario's sig, and clicked it out of curiosity (Thanks for the rate Doc!!). Hadn't seen that technique before and was intrigued.

Working with SAB and PastyPlates in the past was always awkward for me. Can't see shit clearly most of the time and I just felt a bit fumbly.

Moving to agar on plates was a huge jump in being able to see stuff. And moving wedges from plates vs. glad cups was easier for me as well. I'm no expert with a blade and I still felt like I was wrestling with nuggets when transferring to new plates - sometimes it wouldn't come off easy, or I'd fumble around with it...wasn't pretty.

Watching D3monic's technique was helpful. Thought about it, then improvised a solution that I could test quickly with locally available items. Quick trip to a couple stores nearby and ended up at local Target that actually had a 2-pack of metal straws of food grade steel on sale. NOTE: I was a bit particular about the diameter of the holes. Some are small and what I was actually looking for was a smoothie/shake straw (larger than 1/4") - I ended up with one that was about 1/4"OD which seems to be fine in usage.

If you have a steady hand and are practiced and have muscle memory, this may not be a big deal other than aesthetics, which I like a lot.

For me, it was an improvement from the aspect of ease of use and aesthetically it looks pretty cool on a plate. As a noob, it frees me up to concentrate on other aspects of my process and not worry about wrestling with multiple cuts and fishing out and placing a nugget. For me, it's easier to punch, then horizontally stab a disk, carefully lift it out, rotate blade and put it spores down and centered in a new plate.

I would say it's worth looking in to for sure. Big thumbs up from me...YMMV

Here's some backlit plate pr0n and a jar update from 09/03 - jars may get sprung this weekend. Woohoo!





Enjoy.

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
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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: Roger Clemency]
    #26920497 - 09/06/20 05:40 AM (3 years, 5 months ago)

Quote:

Roger Clemency said:
Anyway...nice looking plates most of them. I don’t 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 it’s 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 wouldn’t 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) I’ll transfer once more and then use the plates for Blenderless LI - https://www.shroomery.org/forums/showflat.php/Number/22833314Β ;  It works well and gives you about 10 day colonization of qt jars.




Hiya, thanks for the comments, much appreciated!

I'm pretty sure this happened during innoc, I had a hard time with manuevering the wedge and I was like...well that's probably fucked. Catching that, and observing it to understand how "not to do it" rather than tossing it immediately to see the behavior over time is valuable as well.

Thanks also for the advice on isolation, by doing I gather more experience and insight, and I'm learning that isolation isn't a 1 transfer thing. I don't know if I named this log correctly..instead of Isolation Road maybe I should have called it Isolation Rabbit Hole. LOL

Like you, I think it's gonna go bad although my noob eye has hope. Personal observations:

  • I think I'm getting a better eye for stuff and there's a slight difference in color - myc is a pure-ish sort of white, there's a texture and "look" to it - grey or finer wispy stuff tends to be no bueno.

  • Sweaty jars - not sure if pictures are detailed enough to depict that, but for me, bad stuff tends to generate more condensation. More heat from shit fighting? who knows...overly sweaty jars=no bueno, it's like pre metabolite action.

    NOOB NOTE: With what I know now, I'd probably toss this and not even bother with it. However, since I'm going to run all the jars in separate shoeboxes and in the interest of "science" I want to see where this goes to develop a better understanding and share that with others. You'll still have to make your own mistakes, but hopefully you'll have something to confirm/validate with :smile:

  • Metabolic action - I'm already noticing more than normal condensation at the bottom and darker/wet grain spots in the jar. I'm pretty sure I'm seeing traces of the dreaded golden/amber hue - the jar looks almost right and the noob in me has hope...but my developing Mush Jedi powers say there is no hope...lol...I'm sharing this because I've hit spots like this where you frantically search the shroomery for hope, but you just gotta let go. Unless you willing to take the time to log and learn from the impending crash and burn.

Thanks again for the comments and sorry for not replying sooner. Just got a bit busy and forgot.

I think I'm at a transitionary point of learning where I've got one foot in the noob world and one foot in the beginner world and hope to share info and observations that may make the transition easier for others.

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 (09/06/20 07:10 AM)


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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]
    #26920558 - 09/06/20 07:02 AM (3 years, 5 months ago)

Update:
After trying D3monic's punch technique, I decided to take X3 to X4 with 3 remaining plates since they grew out so fast.

First part looked something like this:

- Xfer using punch was clean, and seemed pretty fast, I literally punched these 3 days ago...

I had a weird idea. I just PC'd 4 x 1qt jars. I thought, what if I took a couple of samples from the area I just punched and nocc'd with that. If they turn out good..I got a fresh plate that has a better than average chance of having those genetics. Especially if I'm closer to a truly isolated culture.

So in the immortal words of Tom Cruise in Risky Business: "Sometimes, you just gotta say. What the fuck!"

So I did a thing:


Labor Day Weekend Generational Family Photo:


Kids on the right are up on deck for spawning today. Based on observation so far and comments, I'm thinking 2A may be a challenge.

Gonna do another sleeve of plates and just got some LME and contemplating trying that. I don't mind the haze from using Potato Flakes, but clear agar looks sooo sexay. I'll be searching for an LME recipe here and online, but if anyone has a favorite they with good results, please post up :smile:

Sorry for all the posts. Trying to separate them contextually.

Hope the info is useful.

Enjoy.

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 10:02 PM)


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InvisibleDoctor Mario
πŸ„πŸ„πŸ„πŸ„πŸ„
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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]
    #26920624 - 09/06/20 07:52 AM (3 years, 5 months ago)

Still looking good! Im anxious to see how it all fruits out.


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InvisibleWall.E
Bacteria's Bitch
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Registered: 06/05/20
Posts: 2,860
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Re: Trying isolation with agar and petri dishes - thoughts? [Re: Doctor Mario]
    #26920824 - 09/06/20 09:44 AM (3 years, 5 months ago)

Those plates look vigorous to say the least, lol


--------------------
Life’s shit, but I’m loving it



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OfflineWunFunGai
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Re: Trying isolation with agar and petri dishes - thoughts? [Re: Wall.E]
    #26921548 - 09/06/20 04:36 PM (3 years, 5 months ago)

Quote:

Myc_Hunt said:
Those plates look vigorous to say the least, lol




I know right? It's been warm and humid lately, but I believe if it gets too warm it goes from being ideal for myc to being happy time for the bad bad.

Trying to keep reasonable expectations. My understanding is just cuz it's vigorous on the plate doesn't mean it'll be that way in the bed...err tub. :undecided: LOL

Thanks for the kind words.

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
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Re: Trying isolation with agar and petri dishes - thoughts? [Re: WunFunGai]
    #26921610 - 09/06/20 05:20 PM (3 years, 5 months ago)

They look really damn good!


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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]
    #26921707 - 09/06/20 06:19 PM (3 years, 5 months ago)

Greetings Mush Cult, Happy Sunday!

An update follows along with some observations and pondering on switching to another tub size.

UPDATE: last night I did a thing...

Prepping Substrate - Nothing fancy here, just a common CVG mix: 1 Brick (682g weighed), 2 quarts verm, 1 cup organic gypsum.



* Break up the brick to rip up any potential long strands compressed in.
* Pour verm, then gypsum on top
* Bring 4qts of water to a full boil, and carefully pour into bucket and seal overnight and mix with a shovel in the morning.
* Double make sure it's mixed evenly (gypsum likes to clump) and test for field capacity before hitting the tubs...
-- Good ole bucket tek works fine for me.


Prepping to Spawn - some updated pics of the 2 xfer group that was nocc'd up on 08/24.



Note that in 2A our suspicion that there was a hitchhiker seems to be correct. Pics should tell the story, and I've post processed them to emphasize the differences; Sweat, amber liquid, excessive condensation while colonizing for me means something is not going well. I may take this through fruiting as far as I feel comfortable to share how things end up down that path.


Feeling Tubby - I've got leftover 6qt shoeboxes left from initial explorations into mini-dubs that I was going to use and spawn each jar to a separate tub, but came across a slightly different sized 9.7Qt box at Target the other day that was square and had a bit more surface area.

Pics for reference:


Sterilite 1642 6qt "The Standard" - 13 5/8" x 8 1/4" x 4 7/8" = 129"sq base ~548" volume
Target Medium Storage Bin 9.7qt - 11.8" x 11.8" x 5.3" = 139"sq base ~738" volume

Anyone out there use this 97qt size before? I usually do a 1:1.5-2 spawn to sub and was curious if more surface area would be a good or bad thing. Probably gonna just stick to shoebox for now as I can fit 4 boxes side by side on my rack.

Comments, suggestions welcome.

WFG :smile:


Edited by WunFunGai (09/07/20 10:02 PM)


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OfflineGalendula
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Re: Trying isolation with agar and petri dishes - thoughts? [Re: WunFunGai]
    #26921803 - 09/06/20 07:20 PM (3 years, 5 months ago)

Hey loving this thread. Cant wait to see how they fruit. Im a noob currently using agar no pour jars but im seriously considering switching to petri... i wana see what im doing and would love to get all scientific!

Amazing tip with the straws ill definately check that out.

The only issue im having with the petis is that they are one use.. im a bit of a zero waste nerd but itll prolly cut down on waste in the long run if i can stop the contam of my grains... does anyone reuse their petris? Iv read on here glass petris are a hasssleback potato to re sterilise.

Mainly just commenting on this thrwad to say amazing please dont stop posting and thank you πŸ¦–πŸ¦–πŸ¦–


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OfflinePearl
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Registered: 05/28/20
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Last seen: 1 year, 3 months
Re: Trying isolation with agar and petri dishes - thoughts? [Re: Galendula]
    #26922428 - 09/07/20 05:06 AM (3 years, 5 months ago)

@WFG: really enjoying following this, keep it up! Super interested to see how these fruit given the detailing of the whole life cycle. :smile:


@Galendula: I'm the same about waste and have looked into reusing the plastic ones with good success so far. From what I've read, a 24hr soak in a 10% bleach solution kills the vast majority of bad.

What I did as a trial:
1. Toss dead/used agar pucks and wash petris
2. Submerge petris in 10% bleach solution overnight (or longer - I ended up leaving them in for over a week due to timing issues)
3. Prep agar as normal.
4. Soak a paper towel in alcohol to use as a draining pad.
5. Put everything in a SAB.
6. Prep SAB as normal.
7. Move petris from bleach solution to alcohol for a quick dunk, then drain on an alcohol pad.
8. Once all dishes are done, pour as normal.


So far I have the same level of issues as with sleeved dishes, i.e. maybe one dish has a problem every now and then.

The question becomes: is this worth the time and effort to you?


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OfflineWunFunGai
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Re: Trying isolation with agar and petri dishes - thoughts? [Re: Pearl]
    #26923205 - 09/07/20 02:35 PM (3 years, 5 months ago)

Another quick update:

Transfers after 24hrs are already showing signs:


Substrate - Made sure to turn and break up any large coir chunks in the substrate. I like to try and make sure everything is broken up well and pull any long strands of coco fiber that sometimes show up:

* This time I'm using a bit less water and shooting for "near" field capacity based on some teks I've read.

These guys are on deck in a bit:


Enjoy.

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 10:01 PM)


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InvisibleDoctor Mario
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Re: Trying isolation with agar and petri dishes - thoughts? [Re: WunFunGai] * 1
    #26923297 - 09/07/20 03:51 PM (3 years, 5 months ago)

Hell yea bro! Moving along beautifully.


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OfflineGalendula
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Registered: 09/02/20
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Last seen: 2 years, 11 months
Re: Trying isolation with agar and petri dishes - thoughts? [Re: Pearl]
    #26923768 - 09/07/20 08:33 PM (3 years, 5 months ago)

Pearl: awesome suggestion I will definitely give that a go. Sounds not too labour intensive... but as I try I'll figure things out. Great basis to start from thank you!

Will be interesting to transfer to a new plate and a 'washed' plate at the same tie and see the difference
πŸ™πŸ™πŸ™


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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: Galendula]
    #26923913 - 09/07/20 10:01 PM (3 years, 5 months ago)

Labor Day Update: Been a lot of labor going on today, yeesh...multi-tasking at max.

NOTE: This is a big update, I reserve the right to reformat or change up later if I feel like it! You have been warned :smile:

Got busy and worked on spawn from 2nd xfer agar to grain. Jars have all been shook. 2A as suspected was a bit gnarly looking - A quick cocktail before starting was in order though:



Jar 2A, I dub thee, Sketchy McSketcherson...for science!!:


Spawn to Tubs: Using 1qt jar to 1qt of sub on the bottom, and a 1qt layer of straight sub on top, then hydrate top layer by misting for about 20 seconds.


Sketchy McSketcherson Tub and Pics:


Finishing Up:


Bewm: 3 promising tubs, 1 on alert, jars started, and 4th xfer moving along...


And now for something different: Gonna mix it up and switch to D3monic's LME agar recipe to do 20x plates for the next leg of isolation road:


Enjoy.

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 (09/08/20 12:17 AM)


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