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OfflineEdmunter
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Agar and isolates
    #23733169 - 10/13/16 05:27 AM (7 years, 3 months ago)

Posted again due to no answers

So what are your usual stages to get an isolate from spores.  I get confused about how many plates to take and end up with too many and the missus threatens to leave with the cat.

germinate
multi spore grow
clone
?????????


How do you continue on and isolate once you like one.

Do you take a wedge from a culture before using it for a grow?

Grow and take a clone from a shroom?

Use and culture and leave a little behind in the plate?


Once you have a culture u like u can then take load of wedges and keep them in the fridge till you need them?

How long can you keep that up for before you need to slant?


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OfflineNDStepp84
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Re: Agar and isolates [Re: Edmunter]
    #23733204 - 10/13/16 06:02 AM (7 years, 3 months ago)

To get/ look for a good culture, many people will do a few ms grows, select some clones, test said clones then slant them on master slants in the fridge  longer term, agar shorter term. No one hardly shoots for an isolate from ms, could take a shit load of transfers before you even begin to see sectoring then each isolate has to be tested and has the same potential to be crap as it does great. 

Clones aren't isolates, but the genetics are narrowed down and at least you have an idea of what you may be getting. If you wanted to isolate from a clone you could easier and test it.

You could store a slant for a year maybe two if done right.  pull a wedge from the slant to a petri, let it grow out a bit and expand said plate to grow again and make a fresh slant.


--------------------

"I am free, no matter what rules surround me. If I find them tolerable, I tolerate them; if I find them too obnoxious, I break them. I am free because I know that I alone am morally responsible for everything I do.
-Robert A. Heinlein 

:takingnotes: Links and teks:takingnotes:
ND's grow log and discussion
Plant thread


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Invisibleblindingleaf
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Re: Agar and isolates [Re: NDStepp84]
    #23733222 - 10/13/16 06:16 AM (7 years, 3 months ago)

:whathesaid:

u either do

MS grow>clone>clean>test fruit>keep/not keep

MS grow>clone>isolate sectors from clone>test fruit each one>keep/not keep

(above two is grower preference, with arguments on both sides as to which might be better)

MS spore swipe>clean>transfer sectors (many plates)>get a single set of genetics (an isolate)>test fruit>keep/not keep

as far as keeping the isolate/wedge DURING the test grow (cause u don't know if its good or not yet) u can just noc up a new plate when u make ur master OR leave some behind on that donor plate, wrap and cold store, thats preference too.

KEEP LABELING TIGHT!!!  seriously.


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MICROBIAL HUSBANDRY!!!!

The whole is greater than the sum of its parts


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OfflineEdmunter
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Re: Agar and isolates [Re: blindingleaf]
    #23733234 - 10/13/16 06:32 AM (7 years, 3 months ago)

Thats great advice guys.  Thanks you so much 

I left this one too long and it wanted to explode.  Im gunna test a plat.



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OfflineNDStepp84
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Re: Agar and isolates [Re: Edmunter]
    #23733556 - 10/13/16 09:29 AM (7 years, 3 months ago)

Agar pins can also be dropped to new plates, grown out and tested. One of my favorite cultures came from an invitro pin:thumbup:


--------------------

"I am free, no matter what rules surround me. If I find them tolerable, I tolerate them; if I find them too obnoxious, I break them. I am free because I know that I alone am morally responsible for everything I do.
-Robert A. Heinlein 

:takingnotes: Links and teks:takingnotes:
ND's grow log and discussion
Plant thread


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OfflineEdmunter
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Re: Agar and isolates [Re: NDStepp84]
    #23747926 - 10/18/16 04:50 AM (7 years, 3 months ago)

Do you need to clean and invitro clone?


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Offlineenlightenment
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Re: Agar and isolates [Re: Edmunter]
    #23747955 - 10/18/16 05:27 AM (7 years, 3 months ago)

The environment in agar plates is pretty clean if they are sealed with parafilm. The tek you used for your 'plates' should allow you to clone a pin without cleaning the pin. Be fast and as clean as possible with agar and success is on your side.

An invitro pin can produce more than one sector. If that's what you meant. An isolate that forms invitro pins does not produce more than one sector because it is an isolated culture.


Edited by enlightenment (10/18/16 05:36 AM)


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OfflineEdmunter
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Re: Agar and isolates [Re: enlightenment]
    #23748356 - 10/18/16 09:25 AM (7 years, 3 months ago)

Great I took 3.  I will test them later.  Can you take a few wedges from a plate before using it to inoc?


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Offlineenlightenment
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Re: Agar and isolates [Re: Edmunter]
    #23748646 - 10/18/16 11:23 AM (7 years, 3 months ago)

Yes that's possible and recommended if you have an isolate or clone. It's like a backup. It is possible that the jar contaminates and the agar work was all in vain.


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