Huskies said:

I received a wedge, but unfortunately it stayed in my mailbox in freezing temperatures for a few hours at least.

When I saw the wedge, it was black like this. I don't think it is mold, looks more like frozen fruits that have burst. Makes sense it would happen like that on agar as well.

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Is that from me? I sent out some x7x+ wedges that were on black agar (with activated charcoal) in those exact same bag type.

I thought I sent a note to the people I sent it to but it's possible I missed one. 

Even if that's not from me, I bet that's the story. Black agar, plus wedges almost always look squishy like that from going through the mail machines.

Here's one (that grew out a bit, packed the bags about 2 weeks before I took this) in the same bag.


Here's a charcoal plate:

cabin sequestering is pretty simple check out this post it links to the video and the timestamp for the demonstration.

Edit:
I just realized that the CS technique does actually call for and use coverslips. You can see that is what they're actually using in their photos in the paper.

Note the clean squares in the top 4 photos, those are the coverslips.


My confusion came from the video demo that is floating around on here by Paul, where he creates the roofs/coverslips with agar from the same or another plate.

From the photos of CS I've seen here on the forums most people are doing this and not actually using coverslips.

They mention this in the paper

Quote:

Cother and Priest [16] described a method by putting a small amount of contaminated agar culture in a Petri dish and covering a large piece of fresh agar, then placing a second small, fresh agar plug on top of the large agar. Once the mycelium has emerged in the second plug, transfer it onto a new agar plate. However, to handle a lager agar is also a difficult operation which easily brings new contaminations.

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fahtster said:

It’s also pretty early.  This may have been premature.. it just looks like it’s going to scallop but I’ll revisit once it’s grown out more.  But I will be trying just the sequestering too.  I’m not sure I get the glass cover slip though… there’s nutrient agar underneath it.  Why would just a piece of glass stop bacteria, especially if the bacteria is riding along with the myc?  Idk, maybe there’s a mechanism going on I’m just not getting atm.  I’m about done thinking about it for now lol

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Quote:

sage_bd said:
Here is a photo of some bacteria on spores:



the abstract it says this:

Quote:

A handy method to remove bacterial contamination from fungal cultures said:
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. [...] 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.

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So its the agar-agar walls of the cabin that are physically blocking the bacteria from riding along on the myc as the myc penetrates the agar. If I'm understanding the paper correctly.

My assumptions here are that myc is really good at penetrating things since it helps trees share nutrients in the forest etc which is how it gets through the sealed agar cabin, hence sequestering the bacteria in the cabin.

Please anyone feel free to correct me if there's a flaw in my logic here, just trying to learn and spread the good word.

edit: I just scanned the paper a tiny bit more and in section 3.1 found the following

Quote:

paper said:
The M. oryzae hyphae can penetrate into the surrounding medium and bypass the coverslip to grow and form new colonies around it. In contrast, the bacteria are restricted by the solid medium and the coverslip, and, thus, they are retained in the cabins under the coverslip. The fungal hyphae around the coverslip are reinoculated onto new CM plates.

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