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 AgarI'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 GelAnother 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 IngredientsThe 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.
Directions1. 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 TransfersThis 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 NeededThe 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)
Directions1. 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