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Roadtripper
Stranger
Registered: 01/16/18
Posts: 2
Last seen: 6 years, 11 months
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Appropriate sectors for strain isolation (pic)
#24919305 - 01/16/18 08:13 PM (7 years, 1 day ago) |
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Hey all, first off i just wanted to give a shout out to all the members for all of the useful information, especially the trusted cultivators. Ive been trying my hand at strain isolation on agar and after 3 transfers am beginning to have a hard time picking out sectors. In your opinion, from the pic, did i choose the correct sectors and how close to a mono culture am i? Thanks again to everyone thats helped move this hobby in a forward direction. Oh and those specks you see are just undissolved agar.
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bodhisatta 
Smurf real estate agent


Registered: 04/30/13
Posts: 61,915
Loc: Milky way
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Re: Appropriate sectors for strain isolation (pic) [Re: Roadtripper]
#24919314 - 01/16/18 08:16 PM (7 years, 1 day ago) |
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doesn't really matter where you pick from you don't know what you got till you test. as long as you don't pick spots that are obviously slow or whatnot.
and you're probably no where near close to an isolate.
if you want to go the isolate route you'll want to get a dozen or more to test because most of them will probably be nothing special.
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bodhisatta 
Smurf real estate agent


Registered: 04/30/13
Posts: 61,915
Loc: Milky way
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Re: Appropriate sectors for strain isolation (pic) [Re: bodhisatta]
#24919319 - 01/16/18 08:18 PM (7 years, 1 day ago) |
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it's far easier to just take what you have and go to grain right now. grow that out and clone something and use those clone cultures
here's an example of the work involved with doing isolate searching https://www.shroomery.org/forums/showflat.php/Number/24741888/
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mushpunx
Fungus Punk



Registered: 04/20/14
Posts: 13,394
Last seen: 2 months, 7 days
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Re: Appropriate sectors for strain isolation (pic) [Re: Roadtripper]
#24919338 - 01/16/18 08:24 PM (7 years, 1 day ago) |
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I don't really see any obvious sectors in that culture. You need to make more transfers first. It can take up to 10 transfers IME. Right now there are so many strains present in that culture its too hard to pick individual sectors out.

Here are some good photos of sectoring and lines of separation I keep around. The one with the drawing belongs to Stro.
The first photo is a transfer I took from a sector, as you can see I grabbed from more than one because it is still sectoring.
The second photo is about 10 or so transfers away from spore germination. This is the point where I would start transferring from the individual sectors- you can see them pretty obviously. The goal of strain isolation is to have many isolates from one culture to test, not just one.
Once you see real obvious sectoring take a transfer from each sector to a new plate. I aim for at least 10. If you took a transfer from a single sector then that will be an isolate, a monoculture. But frequently some of them will still show sectoring because you grabbed tissue from more than one sector (they can be hard to separate sometimes). In that case just make another (or more) transfer until its an isolate.
Then you will have at least 10 different isolates you will need to label, store masters of in the fridge and test each one, keeping only the best. Typically only one out of ten will be worth keeping.

Here are some old photos of isolates I have
--------------------

Amateur Mycologists United
AMU Q&A
Edited by mushpunx (01/16/18 08:33 PM)
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Roadtripper
Stranger
Registered: 01/16/18
Posts: 2
Last seen: 6 years, 11 months
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Re: Appropriate sectors for strain isolation (pic) [Re: bodhisatta]
#24919346 - 01/16/18 08:27 PM (7 years, 1 day ago) |
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Thanks for that link dude, it's a good read. I bookmarked it. If i go to grain and clone a fruit arent i essentially just starting back at MS Since clones still contain multiple sets of genetics?
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bodhisatta 
Smurf real estate agent


Registered: 04/30/13
Posts: 61,915
Loc: Milky way
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Re: Appropriate sectors for strain isolation (pic) [Re: Roadtripper]
#24919352 - 01/16/18 08:29 PM (7 years, 1 day ago) |
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no because you're weeding out the vast majority of genetics and keeping only the ones that made up the mushroom the clone came from. it's just called a clone at that point. not MS and not an isolate. though some clones are isolates right off the bat.
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