36 replies to this topic

[Saphroziac]

I made a few Malt Extract Agar with Yeast Plates: (2%)(MEAY) in FastFred's Media Cookbook.

 

The receipe is:

20 g extra light malt extract 

2 g yeast 

15-20 g agar 

1 L water [10] 

 

Do you think I should store my agar plates in a 52F environment, or in my house in a plastic box in a 60+ environment?  Or put them in my incubators at 70+

 

I use a few of my spore syringes, and did a lot of grain transfers.

Do you think Pan Cyans would grow on this recipe via spore syringe? Should I have used a recipe with Peptone for Pan Cambo/Cyans?

[Jrotten]

Pans will grow fine, if you store them too cold they can be really wet when you take them out. I’ve gone from storing them in the fridge to saving the sleeve and putting them back in and just keeping them around room temp.

[CatsAndBats]

In my experience, it's wild swings in temperature that cause the most condensation.

 

Healthy vigorous mycelium will grow on virtually any agar recipe IME, and I've made pasta agar, silkworm agar, blood agar, ad infinitum..

 

Stay away from anything highly acidic or with significant fat content, unless you want agar jam.

 

If you want to get the spore syringe onto agar, apply the spore fluid to a sterile swab and streak it, IMHO.

[Saphroziac]

I had to yeet around 10 Agar Plates because they had contams in them. Can anyone tell me what is going on in these plates? All 10 were more or less like this.

I'll post the uncontaminated ones next.

 

Edited by Saphroziac, 29 November 2020 - 04:32 PM.

[Saphroziac]

Here or some pictures of the ones that have not contaminated out yet. I think I am going to try and isolate them further. Into 5-10 dishes each. Some of them like Jalisco have nothing in them even after 5-6 days now, should I toss it?

Question, how do I not get the water droplets to form on the interior of the lid, it makes it hard to take photos. When I pour the agar and then close the lid, the heat creates condensation...so I guess I could wait until it cools down. but then it will solidify.

 

 

Edited by Saphroziac, 29 November 2020 - 04:31 PM.

[PJammer24]

They get wet from the temp swings in your fridge and condensation like the Bat-Meista was just saying... You can get a PID/temp controller for your fridge/freezer and keep those swings to a minimum for better storage conditions.  The same units can be used to regulate temperature when steam pasteurizing, etc.... They are pretty handy lil buggers all round... 

 

https://www.ink-bird...controller.html

 

Some units are more robust than others but you should check them out if you're able!  

[Saphroziac]

Currently I am storing them in my Incubator at around 72F.

[CatsAndBats]

I had to yeet around 10 Agar Plates because they had contams in them. Can anyone tell me what is going on in these plates? All 10 were more or less like this.

I'll post the uncontaminated ones next.

 

 

 

Don't forget, agar is a tool that you can use to clean up cultures. When an agar plate shows contamination points like yours does, one can easily pull good myc and move it to another plate/jar. Unless it's sporulated mold, then mos def YEET them!

 

 

 

I almost spit my beer out when I read that, btw.

 

 

Here or some pictures of the ones that have not contaminated out yet. I think I am going to try and isolate them further. Into 5-10 dishes each. Some of them like Jalisco have nothing in them even after 5-6 days now, should I toss it?

Question, how do I not get the water droplets to form on the interior of the lid, it makes it hard to take photos. When I pour the agar and then close the lid, the heat creates condensation...so I guess I could wait until it cools down. but then it will solidify.

 

 

1.jpg2.jpg3.jpg4.jpg5.jpg6.jpg7.jpg8.jpg10.jpg9.jpg11.jpg

 

What was it? Myc? Spores? The desire to yeet is very strong with you, padawan!

 

 

 

 

Patience!

 

 

Currently I am storing them in my Incubator at around 72F.

 

I just posted an agar plate primer that I found very helpful in my transition from jars to plates. You can find it here:

 

https://mycotopia.ne...54#entry1470936

Attached Thumbnails

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Edited by CatsAndBats, 01 December 2020 - 04:44 PM.

[Kmmfarm]

Another good way to mitigate the condensation is pouring your agar at a cooler temperature and stacking your dishes as you pour them. Letting the dishes solidify in a warmer room and storing them at a constant temperature. You could also wait a bit before putting your parafilm on your dishes to naturally let the condensate evaporate. Good luck keep up the good work.

[Saphroziac]

Latest photos of Contaminated and uncontaminated agar plates. :P

 

Not sure what this is.

 

Seems we have bacteria on the sides, I am going to Isolate into a few other plates today.

 

These two are doing the best it looks like. :D

 

The rest of the plates in my Petri Dish Mountain need a week or two more. 

 

 

 

 

[Saphroziac]

Could anybody recommend how many Agar wedges I should put into 1 quart of grain spawn?

[Auhron]

from my understanding, as much as you can. the more you put in the faster it'll colonize, and the more resistant to contamination it will be.

[Arathu]

My method, I use three.......I reason less cutting and moving of the wedges.....it's what I do anyway.....

 

Plate cut into six wedges.....

 

A

Edited by Arathu, 08 December 2020 - 06:56 PM.

[ElPirana]

Could anybody recommend how many Agar wedges I should put into 1 quart of grain spawn?

Lately I’ve been transferring pea-size or slightly smaller wedges of agar, maybe three pieces per quart jar. So I can inoculate at least 40 quart jars from one agar plate.

I used to use bigger pieces of agar and more pieces per jar for inoculation. I realize my method is probably not the norm.

[Saphroziac]

Hey guys,

 

These plates seem pretty clean, so today I am going to test them out and transfer these agar plates into rye quart jars that have been sitting for about a month. 

 

Also, in a lot of my agar plates I see this stuff. I am not sure if it is the agar settling where I cooled off the hot scalpel, or if it's bacteria. I presume bacteria would get bigger over time... I need to watch more agar videos and learn more about contams.

 

Will post pics of jars soon!

 

 

[Saphroziac]

I scratched some spores off a couple spore prints, and did a transfer today. :) 

[Saphroziac]

Do you guys reuse petri dishes? It's a total hassle cleaning them, organizing, and then even if you spray it down with alcohol you still have a high chance of getting contam tghan using the ones straight out of the bag. You can get like 500 dishes for $!00 or something. Super cheap.

[Auhron]

I tried to re-use plastic petri dishes, after initially discarding the contents (this was done outdoors), I washed in regular dish soap, then soaked in a 10% bleach solution for 10 minutes, then dunked in sterile water to rinse the bleach, then submerged in Peroxide for 2 minutes, they were pulled out of the peroxide in front of my flow hood. 21/25 of these "cleaned" plates exhibited signs of bacterial contamination right on the edges of the dish in the corners, underneath the agar I poured. 4/25 were actually clean. Were I to try again, I would increase the concentration of the bleach solution to 15%, and leave them in there longer. 

 

That being said, I will never try again.

I bought some 60mm high borosilicate glass petri dishes I can just bake in the oven or pressure cook to sterilize. More expensive initially, but less wasteful in the long run,  less expensive too (after a good number of reuses). the amount of time / resources I will save in sterilization is worth the investment in my opinion. I don't like single use plastics - even if they are "recyclable". Even the grow bags bother me a little. I am looking for appropriately sized PP5 containers for fruiting blocks that I can modify/utilize and REUSE instead of grow bags.

[rockyfungus]

Good luck with the boro plates. I broke them all within a year
 

[Auhron]

Sorry to hear that rockyfungus. Hopefully I get to reuse the plates enough before I break them. How did they break? just accidental mishandling or did they break during sterilization cycles?

 

Also sorry for the tangent on your thread Saphroziac.