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InvisibleMycolorado
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Re: Strain Isolations on Agar, Pics and Questions [Re: c10h12n2o]
    #23701620 - 10/02/16 08:54 PM (7 years, 3 months ago)

Yeah, I'm familiar with streaking spores, just hadn't seen anything on streaking tissue...after you mentioned liquid inoculation it got me thinking of diluting hyphae in the same way and streaking it...worth a try anyway.


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OfflinePiezo
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Re: Strain Isolations on Agar, Pics and Questions [Re: Mycolorado]
    #23702294 - 10/03/16 01:50 AM (7 years, 3 months ago)

Thanks for the progress update, you have been very busy.

I think they are talking about spore dilution. I never thought about myc dilution for isolation, maybe you should give a try. I looked at LI from soft agar under microscope and filaments tends to stick to others. Maybe a tenso-active agent could help (glycerol ?) ?


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InvisibleMycolorado
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Re: Strain Isolations on Agar, Pics and Questions [Re: Piezo]
    #23702806 - 10/03/16 08:57 AM (7 years, 3 months ago)

Hey Piezo, hyphae sticking together was my chief concern, the other being that the filaments may break up into pieces upon agitation...so yeah, maybe glycerol or even just soap...Dr. Bronner's sal suds or liquid castile?  The sal suds has fir needle essential oil in it so not sure how that would go.  Any other suggestions?


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Re: Strain Isolations on Agar, Pics and Questions [Re: c10h12n2o]
    #23703889 - 10/03/16 02:28 PM (7 years, 3 months ago)

Quote:

c10h12n2o said:

been through around 2000 plates (working with lots of varieties, clone cultures, etc.), and learned a hell of a lot, been doing an insane amount of work. cannot understate how insane the amount of work is.




No shit. I stopped playing with a handful of cultures/varieties at a time for that reason. Even when you're not isolating, having a shitload of cultures that you need to keep up on is counter productive if you don't have the time for it. There is a difference between keeping up with your cultures and really focusing on them.  I try to have no more than 2 varieties going at the same time, that way I can really take my time and focus on them.


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OfflineKenetic
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Re: Strain Isolations on Agar, Pics and Questions [Re: Supalemonhaze]
    #23703944 - 10/03/16 02:46 PM (7 years, 3 months ago)

Shit, I've got my hands full with one.


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OfflineWonderNads
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Re: Strain Isolations on Agar, Pics and Questions [Re: Kenetic]
    #23741825 - 10/16/16 09:15 AM (7 years, 3 months ago)

C10,  really appreciate your time in all this and sharing with us. This has been such a great thread. Also, greatly appreciate your humble attitude and kindness in your responses.  So nice to see you try to keep things civil,  on topic and douse any flame wars. Keep that crap outta good threads! Anyway, can't wait to see your latest progress!


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Formula: 0


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InvisibleMycolorado
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Re: Strain Isolations on Agar, Pics and Questions [Re: WonderNads] * 1
    #23784928 - 10/30/16 10:22 AM (7 years, 2 months ago)

I was able to take some really tiny transfers from some 4% MEA plates.  I also grabbed a couple pins off the first plate pictured.  All are looking a lot more refined. 




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Re: Strain Isolations on Agar, Pics and Questions [Re: Mycolorado] * 2
    #23805752 - 11/06/16 08:53 AM (7 years, 2 months ago)

Here's one of the pin plates. Very pretty.


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Re: Strain Isolations on Agar, Pics and Questions [Re: Inocuole]
    #23861574 - 11/23/16 06:34 PM (7 years, 2 months ago)

Quote:

Inocuole said:
Quote:

weetsie said:
Quote:

Supalemonhaze said:
Quote:

weetsie said:
Even when you don't dilute spores and start with a big black dot  your first plate you can still get isolates in less than 5 transfers.





I'd like to see you try and pull this off and document it.




I'll take you up on that.

Watch this space.





:standingby:




:bumpthread:

:waitingpatiently:


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OfflineTedTheHighlighter
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Re: Strain Isolations on Agar, Pics and Questions [Re: Mycolorado]
    #23880896 - 11/30/16 08:06 AM (7 years, 1 month ago)

I just re-read this thread to re-absorb all the great content on here.

Quote:

Mycolorado said:
I was able to take some really tiny transfers from some 4% MEA plates.  I also grabbed a couple pins off the first plate pictured.  All are looking a lot more refined. 






Are these from multispore? If so, what transfer are these on?


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InvisibleMycolorado
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Re: Strain Isolations on Agar, Pics and Questions [Re: TedTheHighlighter]
    #23881173 - 11/30/16 10:35 AM (7 years, 1 month ago)

Hey Ted,

Yeah, MS. These were somewhere in the 10-15 transfer range.


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Re: Strain Isolations on Agar, Pics and Questions [Re: Mycolorado]
    #23881390 - 11/30/16 11:58 AM (7 years, 1 month ago)

Quote:

Mycolorado said:
Here's one of the pin plates. Very pretty.




By "pin plate" do you mean this is a clone?


--------------------
Alice asked the Cheshire Cat, who was sitting in a tree, “What road do I take?”

The cat asked, “Where do you want to go?”

“I don’t know,” Alice answered.

“Then,” said the cat, “it really doesn’t matter, does it?”


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InvisibleMycolorado
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Re: Strain Isolations on Agar, Pics and Questions [Re: TedTheHighlighter] * 1
    #23881432 - 11/30/16 12:13 PM (7 years, 1 month ago)

Yeah, it was a pin taken from this plate:


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Edited by Mycolorado (11/30/16 12:14 PM)


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OfflinecronicrFacebook
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Re: Strain Isolations on Agar, Pics and Questions [Re: Mycolorado] * 1
    #23881436 - 11/30/16 12:15 PM (7 years, 1 month ago)

Nice work


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OfflineKenetic
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Re: Strain Isolations on Agar, Pics and Questions [Re: Mycolorado]
    #23881830 - 11/30/16 02:16 PM (7 years, 1 month ago)

Quote:

Mycolorado said:
Yeah, it was a pin taken from this plate:





Thats a beautiful pic, belongs in a science book


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Todo Cambia
   

               
                                                :cookiemonster::elmo:



DMT said: Everyone know's me, they just don't know it yet


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OfflineTedTheHighlighter
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Re: Strain Isolations on Agar, Pics and Questions [Re: Kenetic] * 1
    #23882032 - 11/30/16 03:19 PM (7 years, 1 month ago)

Wow, that's awesome.
It looks like its beneficial to clone first if you plan on isolating. Starting the isolating process with cloned mycelium looks like it cuts down a lot of the work


--------------------
Alice asked the Cheshire Cat, who was sitting in a tree, “What road do I take?”

The cat asked, “Where do you want to go?”

“I don’t know,” Alice answered.

“Then,” said the cat, “it really doesn’t matter, does it?”


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OfflineKenetic
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Re: Strain Isolations on Agar, Pics and Questions [Re: TedTheHighlighter]
    #23882082 - 11/30/16 03:31 PM (7 years, 1 month ago)

I agree.  When I first started agar (not too long ago), I had all these dilusions about isolating monocultures and creating a "superbeast", then I started actually working and discovered how hard it is to even isolate a dikariotic strain.  We're talking about a lot of transfers with normal technique


--------------------
Todo Cambia
   

               
                                                :cookiemonster::elmo:



DMT said: Everyone know's me, they just don't know it yet


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InvisiblebodhisattaMDiscordReddit
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Re: Strain Isolations on Agar, Pics and Questions [Re: Kenetic]
    #23882895 - 11/30/16 08:03 PM (7 years, 1 month ago)

was going to send c10 something but looks like he flew the coop.


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OfflineTedTheHighlighter
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Re: Strain Isolations on Agar, Pics and Questions [Re: bodhisatta]
    #23883067 - 11/30/16 09:18 PM (7 years, 1 month ago)

Oh no, you might be right. That's depressing, I want to see how those isolates turn out


--------------------
Alice asked the Cheshire Cat, who was sitting in a tree, “What road do I take?”

The cat asked, “Where do you want to go?”

“I don’t know,” Alice answered.

“Then,” said the cat, “it really doesn’t matter, does it?”


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Offlinepndmnm
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Re: Strain Isolations on Agar, Pics and Questions [Re: TedTheHighlighter]
    #24926454 - 01/19/18 06:54 PM (6 years, 11 days ago)

Reviving the dead for keepsies.

Some differing opinions by those who mostly agree.

Some food for thought as I mindlessly transfer things for no reason except for that I have nothing better to do.

Thanks to everyone that posted.


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Glove box vs. Still Air Box (SAB)



Edited by pndmnm (01/19/18 06:55 PM)


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