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oytothevey said:
If I clone a mushroom from a scraped off bit of its innards on agar, will that then (assuming no contaminants) be an isolated strain? No, it will simply be a slimmed down gene pool. Or will it require agar plate transfers for strain isolation? Correct. Many, many transfers until isolation. Talking 50+.

The cloned mushroom assuming it is an isolated strain using the above scraping off a bit of its innards and wiping in a agar plate method, will it then grow mushrooms identical to the original cloned mushroom? Yes and no. Cloning mushrooms doesn't work how cloning plants works, it's more like humans having babies. Two 6' tall, blonde hair, blue eyed parents COULD have children that are also 6', blonde hair, blue eyed, but it's not a guarantee. Percentage chance is high, but it's not absolute.

Could I then clone those mushrooms for a eternal supply of that specific mushroom strain? Yes and no. You wouldn't want to clone a clone, then clone that clone, etc ad infinitum because each successive clone slims down the gene pool even further. Some of that gene pool includes vigor/growth rate. Multiple clones of clones will eventually slow down, they will start to mutate again and not produce similar looking fruits. That's (a quick and dirty overview of) senescence. You'd want to keep the colony alive on agar and expand through spawn expansion. G2G, LC, etc.

How long can you transfer from a master slant? What do you do when it starts to dry up or run out of mycelium? From what I've read you cant keep transferring from a master slant to make more and then transferring them to make more or something bad happens with the genetics...? The mycelium won't run out, but the slant drying out is a concern. Properly made slants in culture tubes last years. An agar plate poured thick will last up to a year. Transferring A2A will effectively never reach senescence. Senescence is really only a concern when you're expanding spawn too far. If you make one quart jar of spawn, and G2G that to 12 more jars, then G2G those to 12 more each, and so on, THAT'S how you'll reach senescence. The first couple transfers will be fine, the later transfers will start to slow down and break down. I don't have a good reference on how much you can stretch it though.

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Sounds like you've done a lot of research though! You're on a good track my friend

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creamcorn said:
working from a spore print starts all over again.  two monokaryotic mycelia meet, and "mate" and you've got a fresh set of genetics.

using mycelium to inoculate more mycelium (as in an agar transfer, a gran to grain transfer, knocking up an LC from an LC, "cloning a clone") etc is the generation thing, and can be done a couple of times but when you overdo it you can run into problems.

we're not talking about generations in the traditional sense, where you think of kids and parents and grandparents and such... its more or less how many generations we're keeping the SAME set of genetics alive.

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AyePlus said:
Ideal circumstance
Master plate gets made into master slant, at the same time 2nd gen master plate is made from the master plate to be used for inoculating and the master plate and slant go into cold storage. Once the 2nd gen master is gone a new plate can be made from the og master until that one is gone, then at that point you make a new master plate from the slant and continue on like that till the slant is almost gone then you make a new slant from the last master plate.

Keep the genetics as young as possible. Cold storage slows the aging process almost to a stop but after ~ 5 years slants should be plated and grown out and re-slanted, plates redone every 4 months or so.

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https://www.shroomery.org/forums/showflat.php/Number/8468463/fpart/all/vc/1
RR's notes on cultivation:
*It was ANNO that pointed out - young vigorous mycelium does better than old. However, I have gone out 5 G2G transfer generations - without ill effect. Genetics plays a part in it. In nature - mycelium is designed to grow out & fruit in one season. You can FOOL with Mother Nature - but extending it to far out - usually brings on some mutation. G2G GENERATIONS

Some good advice I got for my post that was similar to this a while ago and RR's notes are a great read if you get the time