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Kenetic
Nam Sayin



Registered: 08/24/14
Posts: 4,389
Loc: I don't believe in land
Last seen: 5 years, 3 months
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Isolate dilution method
#23563633 - 08/21/16 05:50 PM (7 years, 5 months ago) |
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Has anyone tried to isolate from a spore solution that was serially diluted to get a small amount of germinating spores?
Let' say you take a small amount of spores from a print and make a spore solution with a weighed amount of water, suck up one mm of spore solution and drop it in a vial with 100 mm of water, do the same for another 100 mm of water, and keep doing this until you get a really diluted solution. Then swipe on agar like you were streaking bacteria.
You could get some strong mono mycelium and put it on a dish with another one and breed a strain? If that's how it works.
Seems like a lot less work than a million transfers, but still a lot of work.
I know cloning isolates are a lot more practical but I'd be more than happy just growing with agar for a while.
-------------------- Todo Cambia    DMT said: Everyone know's me, they just don't know it yet
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bodhisatta 
Smurf real estate agent


Registered: 04/30/13
Posts: 61,889
Loc: Milky way
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Re: Isolate dilution method [Re: Kenetic]
#23563678 - 08/21/16 06:04 PM (7 years, 5 months ago) |
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I've tried to get monokaryotic growth a few times.
you really have to dilute them down. I would try 1:10x3 for a 1:1000 so 1ml to 9ml then 1ml of that to 9 then again I'm sure you know how that works already
I would also try 1:100 (what I did)


which produced these.
https://www.shroomery.org/forums/showflat.php/Number/22753388#22753388
more about it there
I could have picked them up earlier off the plate but it seems there was just that many spores that by the time I even saw growth they had formed dikaryotic mycelium.
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Kenetic
Nam Sayin



Registered: 08/24/14
Posts: 4,389
Loc: I don't believe in land
Last seen: 5 years, 3 months
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Re: Isolate dilution method [Re: bodhisatta]
#23563743 - 08/21/16 06:30 PM (7 years, 5 months ago) |
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I checked out the your post, pretty cool. Didn't notice it was 6 months old though. Sorry to resurrect it.
Anyway I came across a post by RR that is pretty useful and what I'll be doing:
Quote:
RogerRabbit said:
It's better to do a dilution as Terry suggested above, and then allow the diluted spores to germinate into hyphae and grow just enough to combine and form dikaryons. Isolate each colony into new dishes as soon as the dikaryons appear and grow each one individually to fruiting stage, rather than allowing them to combine into a single organism on the original plate(s). RR
-------------------- Todo Cambia    DMT said: Everyone know's me, they just don't know it yet
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