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InvisibleD3_Myc
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Posts: 4,399
Loc: Year Zero
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D3's Perfect transfers and agar TEK. * 65
    #26628234 - 04/25/20 09:35 PM (3 years, 8 months ago)

Perfect Transfers and Agar TEK


I'm frequently asked how I get my agar so clear, the colors and how I get such perfect transfers. The following below is a detailed explanation of the process. There's many ways to achieve similar results but this what works for me. Feel free to experiment with different recipes and nutrient levels.



Tools Required






Petri Dishes




By far Polystyrene and Glass will give the best clarity. I use polypropylene for streaks or other circumstances where I don't expect to show off samples. Nothing beats the cheap disposable dishes though. I've tried no pours, deli cups, glad mini rounds.. none where for me. The particular brand I'm using right now is Celltreat. I'm open to others suggestions on brands. Right now a case is averaging me around $100 for 500 count.

The Agar


I've tried several brands of agar agar and premade mixes. Most left fibrous bits in my petris detracting from the overall appearance. Ultimately I have found that this brand gives consistent results



For nutrients i'm using Maltoferm light malt extract it disolves clean and clear.



For coloring, Misty turned me onto this brand. Wilton Neon Gel it has given best results and color doesn't break down in the PC. But after continued testing and anecdotal evidence it has been found that the red and purple dyes contain red#3 or erythrosin which is naturally fungicidal. It’s caused abnormal, stunted or sporadic growth and my recommendation to avoid those two colors or any other dyes that contain red#3 I’ll be doing a side by side comparison and updating a link here for future users. (Updated 10-22-20) Here's the current link on the experiment thread



Another great brand is Chef Master Neon Gel


Another option is pouring black plates using this Charcoal Powder approximately 1g per 100ml but honestly I don't even measure.

9-15-21 New link for charcoal



Ingredients


The following recipe will produce about 40 petri dishes. You can reduce by half if pouring only one sleeve of 20. I've had best results so far with this amount of nutrients, feel free to experiment with more or less levels of LME. Lower the nutrients the more likely you will get rhizomorphic growth, higher nutes more tomentos. Of course genetics and other environmental conditions come into play as well.

  • 20g agar agar
  • 16g LME
  • 1000ml distilled water


Edit 8-4-21

To switch things up can also add 1g nutritional yeast to the above recipe. I like to pour plates of each and send transfers to both medias.

Note: when using nutritional yeast, agar will become cloudy. I recommend using the charcoal powder for yeast plates.

Directions


1. Add distilled water to pot and begin heating to light boil.
2. While water is heating up, measure out dry ingredients.
3. Once water is beginning to lightly boil, whisk in dry ingredients and reduce heat to prevent boil over.
4. If you have a stick blender, blend for 30-45 seconds thoroughly.



5. Continue to lightly boil agar for 1-2 minutes stirring to prevent foaming and boil over.
6. Add food coloring to media bottles. Personally I use 3 bottles for a 1000ml batch. 3 drops food coloring or 1 drop per 100ml per media bottle. Have fun, experiment with mixing colors.
7. Funnel agar mix into media bottles distributing evenly among bottles. Swirl to distribute food coloring, cap loosely and cover top with foil if you desire. After a few years I’ve pretty much eliminated using foil for most things.



Edit 8-4-21

8. Add enough water to PC for a 20 minute run. For fairly cheap you can get a raised rack for your PC keeps your jars and media bottles out the water. Fill PC to just under or touching the rack.



9. Follow the manufacturer instruction for purging and bringing PC up to a boil prior to adding the weight. I use the modified weight with 2 quarters tapped to it. This reduces rattle and allows the pressure cooker to reach pressures above 15psi.
10. Bring the PC up to 15-17psi and set timer for 20 minutes. Continue to monitor pressure don't allow it to exceed 17psi for safety sake.
11. Once time is up, turn off heat and allow pressure to release naturally, use this time to shower, prep SAB if using one, wipe down your prep surfaces ect.
12. In pot large enough to hold media bottles, heat several inches of water to 140f (60c) or if you own a sous vide this is even better.
13. Carefully remove media bottles from PC, place into pot of water ( this serves two purposes, one it helps cool the bottles down to handling temps and helps keep bottles warm to prevent from setting up while pouring other bottles) move your pot and media bottles to where you will be performing pours.
14. Using a laser temp probe measure the media bottles temperature. I begin pouring once temperature reaches 135-140f (57-60c)

Once all plates are poured leave unwrapped in SAB with arm holes covered for several days or several hours in front of flowhood to allow condensation to subside.

 

The end result is a crystal clear plate with no condensation




Taking Transfers


This method of taking transfers can be performed within a SAB or in front of a flow hood. For obvious reasons a flow hood is much easier to work in front of.



Tools Needed


The main focus of this method utilizes a punch or stainless steel straw to act as a cookie cutter making perfectly round holes in your agar for transfer. The method has been floating around on the groups for some time now. You can make one these out of brass tubes, metal straws what ever. You're only limited by your imagination. I’ve been using a custom punch made with brass tubing and has an acrylic handle.



  • Leather hole punch
  • Flowhood or SAB
  • Scalpel
  • Torch or alcohol burner
  • Parafilm or cling wrap... what ever you prefer wrapping your plates with
  • Petri dishes transferring from.
  • Poured petri dishes for transferring to.
  • Spare dish for cooling heated tools ( I use a ugly dish, one that had sediment or perhaps agar started to gel toward end of pours)



Directions


1. Select the plates you will be transferring from.
2. Prep you work surfaces as usual, wipe down with iso ect.
3. Flame sterilize your scalpel and place into a container blade up.
4. Flame sterilize your punch.You will not be able to get the steel red hot without burning yourself and not being able to handle the tool. Just gently flame the tip and slot opening for a few good seconds like shown in video above.
5. Cool the leather punch / transfer tool in the spare petri dish by gently depressing into agar several times until it stops sizzling. If using brass tubing, ss straw ect this will be not necessary as the metal cools fairly quickly on its own.
6. Use the punch to cut perfect lil circles into the leading edge of your myc





7. Using your previously flame sterilized scalpel, gently poke into the side of your transfer disc and lift. This may take a little practice but it's extremely easy once you have done it a couple times. i recommend practicing on a blank petri until you get the hang of it.







By gently pressing the disc onto the recieving plate it should stick and allow you to slide your scalpel out. Sometimes if your scalpel slipped too far into the disc you may have a little difficulty getting it free.

You're left with a perfect little transfer disc in the center of your agar.



The end result after a few transfers are beautifully symmetrical dishes.





Hope this little tutorial helps you elevate your agar and transfer game. I'll update periodically with better videos and pictures.

Thanks, D3


Edited by D3_Myc (10/22/21 08:03 PM)


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InvisibleMurphSmurf
Float On
I'm a teapot User Gallery


Registered: 02/08/18
Posts: 68
Re: D3's Perfect transfers and agar TEK. [Re: mongoosed] * 5
    #27800540 - 05/31/22 03:45 PM (1 year, 7 months ago)

D3's post is what got me out of the no-pours & into "traditional style" agar. Literally bought everything needed off this post. Poured my first 500 plates now, was rough initially, but gradually got better.





been playing w/ millet flour. Myc seems to like it.



Will always be very appreciative of this post.


--------------------
Long-time lurker - love this site - love to learn - love agar - MMFam


Edited by MurphSmurf (05/31/22 03:46 PM)


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OfflineMeiSucks
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Re: D3's Perfect transfers and agar TEK. [Re: dowodenum] * 4
    #28216476 - 03/05/23 07:36 PM (10 months, 18 days ago)

Used this Tek for my first time pouring :boombox2:





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InvisibleGrumio


Registered: 10/18/19
Posts: 126
Re: D3's Perfect transfers and agar TEK. [Re: ouuwee] * 3
    #26655438 - 05/07/20 03:55 PM (3 years, 8 months ago)

I just use metal drinking straws now after getting annoyed waiting for punches to cool down.

They are thin wall stainless steel so ideal for this use and your hands are further away too, even better if you use the bent straws as well to save your hand hovering above.



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OfflineSockadin
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Re: D3's Perfect transfers and agar TEK. [Re: PMBastian] * 3
    #26676296 - 05/17/20 05:47 AM (3 years, 8 months ago)

Demonic punch action...



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OfflineSockadin
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Re: D3's Perfect transfers and agar TEK. [Re: D3_Myc] * 2
    #26628238 - 04/25/20 09:36 PM (3 years, 8 months ago)

YES!

Great TEK. Well written and lots of detail, nice one!


Edited by Sockadin (04/25/20 09:51 PM)


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OfflinecronicrFacebook
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Re: D3's Perfect transfers and agar TEK. [Re: Crackatoa] * 2
    #26629591 - 04/26/20 11:26 AM (3 years, 8 months ago)

always a fan of your work:cheers:


--------------------

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I'm tired do me a favor


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OfflineFrankRhizo
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Re: D3's Perfect transfers and agar TEK. [Re: Mr Solo Dolo] * 2
    #26652181 - 05/06/20 04:52 AM (3 years, 8 months ago)

I'm going to look around my house get creative and find things to use. My first idea is a dab straw.


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Invisibleeatyualive
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Re: D3's Perfect transfers and agar TEK. [Re: D3_Myc] * 2
    #26697643 - 05/26/20 06:03 PM (3 years, 7 months ago)



--------------------
EAT GETS SHIT DONE


:flame::chief:JOIN THE POW WOW:chief::flame:


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OfflineSockadin
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Re: D3's Perfect transfers and agar TEK. [Re: D3_Myc] * 2
    #26700077 - 05/27/20 06:16 PM (3 years, 7 months ago)

D3monic you started a movement man! How awesome is this TEK!


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InvisiblebodhisattaMDiscordReddit
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Re: D3's Perfect transfers and agar TEK. [Re: sandman420] * 2
    #26762468 - 06/21/20 08:33 AM (3 years, 7 months ago)

Quote:

sandman420 said:
This is great stuff I will be trying this soon. What fantastic pictures too. You can really see the difference in clarity in good old fashioned PS petri dishes versus PP dishes that eats is using. It's night and day truly. Disposables are king, shame about the waste issue.



I think of waste from a use standpoint. The dishes lead to plenty good. But something like a water bottle or straw are pretty useless. Most people willingly waste plenty of plastic and water all day but throw a fit at it in the mycology perspective. I don't get it.


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InvisibleBenson
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Registered: 09/29/20
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Re: D3's Perfect transfers and agar TEK. [Re: Huskies] * 2
    #27252337 - 03/14/21 10:39 AM (2 years, 10 months ago)

Probably not the first to try this but I put a lil bend in my stainless steel straw and now my disgusting hands never have to hover over the plate. Also I found it cuts down my SAB hand movements significantly :thumbup:




No healthy plates were harmed in the making of this video :cool:


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InvisiblePsillySeeEms
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Re: D3's Perfect transfers and agar TEK. [Re: Irishmule169] * 2
    #28174419 - 02/06/23 06:01 AM (11 months, 15 days ago)



:loveheart:


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InvisiblePsillySeeEms
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Re: D3's Perfect transfers and agar TEK. [Re: PsillySeeEms] * 2
    #28202208 - 02/24/23 09:43 AM (10 months, 27 days ago)





Anyone checked out these angled dental tissue punch kits on ebay?

$13 for a 4 pack, which includes sizes from 4-8mm.

Or, $6-8 a piece. Free shipping.


Edited by PsillySeeEms (02/24/23 09:46 AM)


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Invisibledowodenum
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Re: D3's Perfect transfers and agar TEK. [Re: PsillySeeEms] * 2
    #28202883 - 02/24/23 04:28 PM (10 months, 27 days ago)

Nice, those look good.

I just went the straw route. I think the size is about 8mm?



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Invisiblewoofwoof
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Re: D3's Perfect transfers and agar TEK. [Re: Sockadin] * 1
    #26628260 - 04/25/20 09:48 PM (3 years, 8 months ago)

Nice job!


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InvisibleSubnet Mask
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Re: D3's Perfect transfers and agar TEK. [Re: Sockadin] * 1
    #26628276 - 04/25/20 09:56 PM (3 years, 8 months ago)

:rockon:


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Invisiblemushhead
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Re: D3's Perfect transfers and agar TEK. [Re: Subnet Mask] * 1
    #26628283 - 04/25/20 09:58 PM (3 years, 8 months ago)

I've been waiting for this specific write up!


--------------------
Meditation Principles

Silence: Giving you room to listen / Stillness: Giving you room to feel / Spaciousness: Just giving you room


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Invisiblegizmo1
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Re: D3's Perfect transfers and agar TEK. [Re: D3_Myc] * 1
    #26628518 - 04/26/20 12:17 AM (3 years, 8 months ago)

That's some really good looking growth πŸ‘Œ
D you havent had any problems with contams dropping of your hand onto the open plate or the piece being transferred?
I get contams everytime I accidentally pass my gloved hand over a plate. Especially when I'm pouring. I always know which sleeve is going to have a bad plate in it. I see the pic where you are cutting the disk and it scares the shit outta me lmao. I mean it seems to be working good for you but it makes me uncomfortable for some reason.


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Trade List
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InvisibleHobbit GDF
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Re: D3's Perfect transfers and agar TEK. [Re: D3_Myc] * 1
    #26629336 - 04/26/20 09:43 AM (3 years, 8 months ago)

Grateful for your artist approach.


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