PuffM4gicDr4gon said:

If I had to make a guess I would say it's the bacteria that faht talks about

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Twigg said:

Hello all!

I've had a plate of pan Jamaica in the back of my fridge for the past 7 years.



Does it look worth trying to revive?

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fahtster said:

They look good to me.  I’d still do aerial myc transfers and keep that going.  At best, it’ll clean the cultures more each time you do one and at worst, it’ll maintain the cleanliness you have now.  If you do agar wedge transfers and there’s bacteria on the agar wedge, it’ll just compound the problem with each transfer until it’s eventually an unusable culture, IME anyway. 

It’s a numbers game.. if there’s biofilm on the agar wedge, it’s an established colony and very quickly will migrate to the new transfer plate.  Aerial myc transfers may have free floating bacteria on it but the amount of mycelium out numbers those free floating Bacteria.  The myc will establish itself on the new transfer plate first (depending on the aggressiveness of the bacteria) and outrun the bacteria.  If you keep doing this multiple times, the aerial myc should have less and less free floating bacteria on it as it gets further and further away from newly established bacterial colonies on the new plates

Not saying don’t send those plates, just that you should take an aerial transfer from the leading edge before you send them to keep cleaning the culture up

It’s all about numbers with pan.. gotta do a shit load of clones and then a shit load of transfers.  Eventually, you’ll get to a point where you either say, “this looks really good” and send it or you’ll say, “I’ve done 5 aerial transfers and this is as good as this culture is going to get” and you send it.  There’s only one way to test a culture and that’s to grow it out.  Ideally, you’ll want to keep track of each culture and what happens when you send it.  That way, you can either confirm it’s cleanliness or know there’s more work to do.

Keeping your old early transfer plates around is a good idea too.. a fully grown Out plate will tell you more about the state that plate is actually in as far as how clean it was.  You want to watch out for leading edges that crash out or discoloration under the myc.  Early on, a plate may not show discoloration but as the plate grows out, the biofilm builds and that discoloration is more pronounced.  So keep your old plates around for a while and check on them as they grow out.  It all helps inform how you should move forward with the culture.

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Lo-Z said:

Hahahaha you guys are funny.

Faht does talk about that biofilm bacteria that's deeply embedded, almost symbiotically. So far trenching and aerial transfer to a trench seems to work the best. It seems to like to propagate through agar. if there is a gap in the agar, it cant cross. That and Cabins. I've been running cabins and then transferring straight to a trench if there is any contact with agar to agar the biofilm seems to pass through.

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Anomalocaris said:

how does not take aerial myc, just run the scalpel through it and it will usually take a piece?

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Anomalocaris said:

Lo-Z said:

Hahahaha you guys are funny.

Faht does talk about that biofilm bacteria that's deeply embedded, almost symbiotically. So far trenching and aerial transfer to a trench seems to work the best. It seems to like to propagate through agar. if there is a gap in the agar, it cant cross. That and Cabins. I've been running cabins and then transferring straight to a trench if there is any contact with agar to agar the biofilm seems to pass through.

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wow a cabin AND then a trench?

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Yes, what I did was put it through a cabin


Then transfer a wedge straight to a trench


Or another Cabin



Mycelium Olympics

What I've found is exactly what Faht said... If there is a biofilm present (Even if you cant see it at the moment) it will hitch a ride on the agar itself. If you take an aerial transfer like the video Kloakk made, it's still possible that there could be some bacteria / biofilm coupled with the myc, BUT, not as much as there would be with the piece of agar.

Example This is a TTBVI plate that has a biofilm deeply embedded. But we couldn't see it yet. It looks really healthy.


I took an aerial transfer and then days later the biofilm showed up on the original plate.


I took that aerial transfer and sent it straight to a trench


It seemed good for about a week and then it got the biofilm too! You'll  have to zoom in and look close at the small island where the transfer was placed there's a small circle of discoloration.


I think the trench works great for this biofilm becasue looking at this other plate that had a whole wedge transferred to it, we can see the biofilm ring and it doesn't propagate over the trench. though, i'm still waitting to see the results of these transfers becasue the biofilm doesn't show up for about 1-2 weeks on the plate if it is there.



This is what a really deep ring looks like


This plate was made on 10/25
The picture was taken on 11/05 and looked great.


and the biofilm didnt show up until 11/08

Lo-Z said:

Kloakk made a video on how to do it.

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Not sure what is happening there and it also seems to be in a lower quality if watched in a browser as opposed to the youtube app on phones.

This should work