KlarnEvil9 said:

I messed around yesterday doing kitchen stuff and didn't get to that plin... now I have spores on my plate!

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Good catch.

Plins are the easiest way to clone, but you have to get them before they drop spores.

To clone them I cut off the head and the bottom.

Pic tax, the plin that I cloned that grew the shrooms that eventually had the tall mutant whose spores started x7x+:



After growing about 20 shoeboxes of the strong clone, I got a single much taller mutant, and took spores. The ones I sent out where F8 downline from that (selecting for biggest / fastest growing / more robust):

LightShedder said:

The subsequent transfers will contain the multispore genetics included with the clone... that's the point. So you are permanently ruining the clone material because it has spores on it.

With a regular clone picked from a grow,  it doesn't matter if the mushroom was exposed to sporulation or not because you're taking clean unexposed tissue from the inside, whereas with a plin you're just transferring the entire fruitbody without dissecting it and therefore the spores that are present will make the clone transfer impure with spores and then you're growing spores again.

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wichita_jack said:

i am new to agar. i have now taken two clones to agar using Frank's method, both of which grew out full plates w/o any visible contams. i have been using a SAB, which seems to have worked well for those.

Using the same sterile technique and even at the same time as the second clone (before opening the bag for the clone, just in case it brought contams to the SAB), i also tried to inoculate 4 plates with LC to be sure the LC was clean (two different syringes from two different vendors, fwiw). I did one plate for each LC w/ just a couple/few drops and one plate w/ a couple/few drops and then streaked w/ a loop.

my issue is i have absolutely no growth whatsoever, not even some contams, in the 4 plates using LC. all plates, including those from the clones, come from same batch from same vendor. they are ~2:3 agar:LME.

Any ideas what I could be doing wrong?

does all of the liquid in the LC syringe contain inoculant? or is just the part where i can see the myc floating around?

appreciate any help! i would share pics but its just four blank plates.

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LadysKnight said:

KlarnEvil9 said:

I must be seeing the folly of my slack-ass flaming protocol.

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Maybe. But it looks like more than one plate has it? If so, what's your donor plate look like?

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Here are the T2 plates:





You can see how far back the transfers were. But the edges have gotten more rhizo as it spread out to the edges.

I do have 1 decent looking T3 (I think):


And a plin transfer from before the bigger one formed:

Here's the big plin today:




I also have a couple of Josex poke inoculated BRF pucks, but I used jelly jars with the bumpy texture, so I can't see shit yet. Fingers crossed. Thanks to everyone, your expertise is the only way I'm going to have success with this culture.

LadysKnight said:

wichita_jack said:

i am new to agar. i have now taken two clones to agar using Frank's method, both of which grew out full plates w/o any visible contams. i have been using a SAB, which seems to have worked well for those.

Using the same sterile technique and even at the same time as the second clone (before opening the bag for the clone, just in case it brought contams to the SAB), i also tried to inoculate 4 plates with LC to be sure the LC was clean (two different syringes from two different vendors, fwiw). I did one plate for each LC w/ just a couple/few drops and one plate w/ a couple/few drops and then streaked w/ a loop.

my issue is i have absolutely no growth whatsoever, not even some contams, in the 4 plates using LC. all plates, including those from the clones, come from same batch from same vendor. they are ~2:3 agar:LME.

Any ideas what I could be doing wrong?

does all of the liquid in the LC syringe contain inoculant? or is just the part where i can see the myc floating around?

appreciate any help! i would share pics but its just four blank plates.

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Typically, if you see no growth on agar, it's too dry. You mention 2:3 agar:lme, you don't mention the water content, and may not know, but those ratios are backwards for good growth, too much sugar, should be closer to 500ml water/10g agar/6g lme... all the way up to even ingredients, but never more sugar than agar. You really should make your own to know what's in it and how sterile they are. Plus they are 10x cheaper.

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