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Agar blues


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#21 Arathu

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Posted 27 July 2021 - 06:38 PM

I'm sorry I didn't see your question til now......sad......my bad..

 

A broad spectrum of what you can get.....most will not stand the heat of sterilization though......so they have to be added later

 

Gentamycin I've used and also fish and other pet antibiotics too.....

 

A


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#22 Salty117

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Posted 27 July 2021 - 06:47 PM

Quite alright Arathu, I appreciate the info  :smile: I look forward to crafting some antibiotic agar to help me salvage cultures.

 

Thank you


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#23 CatsAndBats

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Posted 28 July 2021 - 12:12 AM

Just say no to antibiotics!

 

Try this instead:

 

Contamination control and removal are very important technical aspects of microbiological research. Bacterial contamination is very common in fungal cultures. Currently, the commonly used approach for inhibiting bacteria is antibiotic treatment; however, there are drawbacks to using antibiotics, including incomplete removal, limited antibacterial spectra, tendency toward recontamination, effects to fungal strains, and potential risks to the environment. Therefore, in the present work, we developed a new method for bacterial removal from fungi cultured on solid medium, the Cabin-Sequestering (CS) method, based on the different culture characteristics between fungi and bacteria. First, 3–5 mm round or square holes (the “cabin”) are excavated on a solid medium plate. The fungal strain containing possible bacterial contamination is inoculated into the cabin. The cabin is then covered with a sterilized coverslip, followed by incubation at the appropriate temperature. After 7–10 days of culturing, fungal hyphae grow out along the edge of the coverslip; however, the contaminating bacteria cannot pass through the space formed between the medium and the coverslip and, thus, remain in the cabin. The newly grown fungal hyphae around the coverslip are re-inoculated into fresh culture plates, where they form bacteria-free fungal colonies. The CS method is easy handling, with a short experimental cycle and rare recontamination. When necessary, it can also be used in combination with antibiotics in bacterial removal operations.

 

From: https://journals.plo...al.pone.0224635


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#24 Salty117

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Posted 30 July 2021 - 04:24 PM

Wild Tennessee Cube transfers from PDA to a recipe that my ADD got in the way off, meant to make PDA w/ Yeast and Charcoal but I forgot that I didn't add dextrose until I was in the middle of pouring my cups. Still lookin' good and trucking along, transfers made on the 27th

 

Super Orange - Wild Cubensis from Tennessee PCYA (1)a.jpg

 

Super Orange - Wild Cubensis from Tennessee PCYA (2)a.jpg


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#25 Oldpunk

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Posted 30 July 2021 - 11:06 PM

Those look pretty good Salty. Maybe the sugar isn't so necessary. Or maybe just a minimal amount.

A pinch of charcoal and a pinch of yeast?

Might have to try that some day.
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#26 Salty117

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Posted 31 July 2021 - 06:41 AM

Thanks!  :victorious: I have yet to try germinating straight to Charcoal and yeast agar, so far I've only made transfers to it. I get worried I'll way over-inoculate the plate and make a mess of things considering how dark the charcoal makes the agar.

 

I also realized when looking at Seeker's agar thread that I was also using WAY too much Peptone before when I was still having problems with the recipe which I've cut out for a little while to keep things simple and get things on rails. Idk if that was contributing to the strange gelatinous and clear yet still oddly rhizomorphic growth that would end up stalling or instead reach up instead of out but cutting the too much peptone and dextrose out seemed to get things on track.



#27 Arathu

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Posted 31 July 2021 - 06:52 AM

I have great success with the methods I use, so I think I'll just say YES when necessary.....y'all are free to do what you want.....

 

Don't use anything if you don't have to, transfer away from infections (repeatedly), this is, in my opinion, one of the points of learning to do agar work....

 

Learn to clone and collect spores carefully.....but there will be cases of bacterial infections for certain.....

 

But the antibiotics work like a freaking charm.....these are all tools in the toolbox IMHO..

 

I do like the idea of the well/pocket.....I will have to fiddle with sterilizing slip covers now......damn it......

 

A


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#28 MushLuvR

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Posted 31 July 2021 - 12:15 PM

I keep it simple with AGAR... 10/10g of AGAR/LME per 500ml or 2g each per 100ml if doing small batches of plates.  I do 20 plates with 500 mils consistently(you won't in the beginning trust me).  


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#29 Salty117

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Posted 31 July 2021 - 01:52 PM

Yeah I've been getting better about pouring less and picking smaller bits to transfer from. I gotta keep telling myself "just a grain of rice" or smaller when I'm doing my transfers  :meditate:


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#30 Salty117

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Posted 01 August 2021 - 04:02 PM

Thinkin' tomorrow night for the next set of transfers? Maybe the morning after.. Or should I wait until they're about to crawl up the walls?

 

Tennessee Cube (1).jpg

 

Tennessee Cube (2).jpg

 

Huautla Cube.jpg


Edited by Salty117, 01 August 2021 - 04:03 PM.

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#31 MushLuvR

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Posted 01 August 2021 - 04:13 PM

That's more than enough growth to transfer(or choose) from.  I like the outer edges and ROPEY for Cubes.  


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#32 Salty117

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Posted 01 August 2021 - 04:15 PM

In that case I may make the transfers after I pour today's batch of agar. No sense in shutting the floow hood down if they're ready lol


Edited by Salty117, 01 August 2021 - 04:15 PM.

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#33 Salty117

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Posted 01 August 2021 - 05:39 PM

I keep it simple with AGAR... 10/10g of AGAR/LME per 500ml or 2g each per 100ml if doing small batches of plates.  I do 20 plates with 500 mils consistently(you won't in the beginning trust me).  

 

I almost got there with this pink water agar I whipped up lol. S.O.'s friend came by to visit and see the new home and asked if I could make it pink, so I did. Since the pink BRF agar I poured I realized it makes it difficult to notice lipstick mold but I wasn't about to be a party pooper about it. Managed to pour 18 3.25oz cups.. the last splash got chunky or else I might have gotten 19 lol.

 

Pink Water Agar.jpg

 

228668857_962529220961493_7028761847509035392_n.jpg


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#34 Salty117

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Posted 01 August 2021 - 07:23 PM

Purple PDA w/ .5g yeast and .75g Peptone. this batch is intended for transfers only. Cambodian, JMF, and McKennii spores are going to the pink water agar shortly and when the purple PDA is cooled I'm going to make those transfers from the other cups

 

Purple Agar (2).jpg

 

Purple Agar (1).jpg


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#35 Salty117

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Posted 06 August 2021 - 06:48 PM

Getting closer and closer

 

Wild Tennessee Cube transfers

 

Super Orange - Wild Cubensis from Tennessee PDPYA (1)a.jpg

 

Super Orange - Wild Cubensis from Tennessee PDPYA (2)a.jpg

 

Super Orange - Wild Cubensis from Tennessee PDPYA (4)a.jpg

 

Martinique transfer. Have more but this one is the most visually appealing

 

Martinique Cube - PDPYA(a).jpg


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#36 Salty117

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Posted 09 August 2021 - 07:59 PM

Tennessee cube progress. Beauty shots before getting transfers taken from them.

 

SuperOrange t2a.jpg

 

SuperOrange T2C.jpg

 

SuperOrange t2e.jpg

 

SuperOrange t2f.jpg

 

SuperOrange T2G.jpg

 

SuperOrange T3c.jpg

 

SuperOrange T3G.jpg


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#37 Salty117

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Posted 09 August 2021 - 08:20 PM

Martinique and Huautla shots

 

Martinique T2b.jpg

 

Martinique t2L.jpg

 

Huautla T2F.jpg


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#38 Salty117

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Posted 12 August 2021 - 09:32 AM

Made a bunch of transfers yesterday

 

Transfers 8.11 (1)a.jpg

 

Transfers 8.11 (9)a.jpg

 

Transfers 8.11 (7)a.jpg

 

Getting transferred today will be these fellows

 

Transfers 8.11 (3)a.jpg

 

Transfers 8.11 (8)a.jpg

 

Transfers 8.11 (5)a.jpg

 

Aaaand I am wondering, are these looking alright? Doesn't look like it's behaving like Cube mycelium. These did come from a tray that had a strange brown spot forming at the edge of the plate by the wall of the condiment cup, the transfers were taken from mycelium on the complete opposite side of the plate.

 

Transfers 8.11 (11)a.jpg

 

Transfers 8.11 (6)a.jpg


Edited by Salty117, 12 August 2021 - 09:34 AM.

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#39 MushLuvR

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Posted 12 August 2021 - 12:25 PM

AGAR is so fun to play with, makes you feel like a KID Scientist or something.  Speaking of AGAR, I only have 3 plates left, I need to make a batch(of 20).  


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#40 Salty117

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Posted 13 August 2021 - 01:01 PM

Transfers (2)a.jpg

 

Transfers (4)a.jpg

 

Transfers (5)a.jpg

 

Transfers (6)a.jpg

 

Transfers (12)a.jpg

 

Transfers (9)a.jpg






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