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Zmush
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Registered: 02/11/24
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Help with Agar Plates, Contamination? PHOTOS
#28655718 - 02/11/24 04:09 PM (10 months, 30 days ago) |
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Hello shroom fam. New here..
Got my hands on some spore prints and have transferred them to Agar.
Agar recipe I use 10g Agar 7.5g LME 0.25 Nutritional yeast
I use no pour with these cups https://www.amazon.com/gp/aw/d/B083HXXB34/ref=ox_sc_saved_image_2?smid=A7OZ9T0YXXHC3&psc=1
No Pour PC method (Presto Cooker 22Quart) - I mix my ingredients and let it stir with the magnet holder for 5 min. Let it come to boil in microwave and repeat the process 3x times. - I pour thin - Fingertip tight lids for gas exchange - Tinfoil on each of the lids - don't know if this is something i really have to do. -Vent for 15 min -Pressure cook at 15 PSI for 25 min
Condensation TEK - Once I have vented and cooked it for 25 min. I let it cool down. As soon as it comes down to 0 PSI and the Security lock is open I take out my dishes. Place them on a foam mat with a metal brownie bar on top with boiling hot water. I then cover the brownie pans. I let it stay there over night. I get no condensations with this TEK. Credit- https://www.shroomery.org/forums/showflat.php/Number/27189961/fpart/all
Here are the plates I have sacrificed by opening the lid for getting the best photos. They are at transfer 3 at day 5. As you see, when you place the agar at a different angle under light, the Agar is cloudy and possibly some mold contamination hence the slime?
1 - Upfront photo
 1 - Photo with different angle, you see the cloudy spot
 1 - Nutritional Yeast Sediments - I did not blend it

2 - Photo
 2 - Angle

3
 3A

4
 4A

Here are some other photos of my transfer 3 at day 4. I have not opened the lid. But the Agar is cloudy / slime looking. I can't honestly tell.







This one is Transfer 2 which I have stored in the refrigerator.


These Agar plates are right of the PC once it has solidified. As you see, its cloudy. So no contamination? What could the reason be




Many of my other plates have the similar cloudy/slime Agar. Most of agar plates also come out cloudy right of PC. Are those contamination or may it be due to Nutritional Yeast?
I'm using an expensive organic local AGAR brand, BRIESS golden light LME and local organic Nutritional Yeast.
I place my Agar plates in a clean box at room temperature.
I'm open to feedback. Please help.
Edited by Zmush (02/11/24 05:18 PM)
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CocaineBuffet
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Registered: 08/29/19
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Re: Help with Agar Plates, Contamination? PHOTOS [Re: Zmush]
#28656098 - 02/11/24 08:21 PM (10 months, 30 days ago) |
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Those black plates look sketchy.
Most of those plates look fine. I think I see what you mean by the slime but it doesn't look like the culture is impacted by it. Is it possible that the plate was moved before the agar solidified causing an uneven media?
Some of those open plates don't look like they needed to be sacrificed.
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Zmush
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Registered: 02/11/24
Posts: 49
Last seen: 1 month, 18 days
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Re: Help with Agar Plates, Contamination? PHOTOS [Re: CocaineBuffet]
#28656575 - 02/12/24 10:15 AM (10 months, 29 days ago) |
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I took them out as soon as the cooled down enough to take out of PC and did the condensation TEK as described.
I’ve made a bunch of plates, had to take of the lid to show what I mean.
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Blufuut180
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Registered: 11/14/23
Posts: 187
Loc: Some where else entirely
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Re: Help with Agar Plates, Contamination? PHOTOS [Re: Zmush]
#28656744 - 02/12/24 12:34 PM (10 months, 29 days ago) |
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I have never had luck with no pour teks. Before I had my flowhood I only ever really had success with pouring hot PDA into ketchup cups in a SAB. To be honest the uncolored plates do not look contaminated. I would do a control batch for your next one. Let them sit for a week to see if anything grows, this will show you if your agar tek is the issue or if something else is.
-------------------- Balance is for men who learn to know.
  
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Zmush
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Registered: 02/11/24
Posts: 49
Last seen: 1 month, 18 days
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Re: Help with Agar Plates, Contamination? PHOTOS [Re: Blufuut180]
#28657078 - 02/12/24 04:18 PM (10 months, 29 days ago) |
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Do you guys also have that cloudy agar or is it only me?
Edited by Zmush (02/13/24 01:47 AM)
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