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OfflineMushmix
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Isolating on agar from ms question
    #26355319 - 12/01/19 10:27 AM (4 years, 2 months ago)

I would like to isolate a decent strain from ms syringe to inoculate a slant tube for long term storage.

The proceedure as I understand it (simplified):

One drop from the ms into each compartment of a four compartment petri dish.

Grow till approx 1cm then transfer a sample from each of the four compartments into the four compartments of a new dish and so on... Isolating sectors until there are no more.
Finally transferring the final mono strain into a tube for storage.


To ensure I end up with a decent strain in the end, how many plates of four should I make from the initial ms syringe?

How many plates should be made from those etc?

Essentially im trying to figure out kind of minimum ballpark, what to keep, what to discard, how many plates and isolations it takes to get a decent master strain using four part plates?

Do you take only the best sectors from each of the four previous compartments and only transfer those to the new plates, discarding the rest?

Storage space storage space lol.


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InvisiblebodhisattaMDiscordReddit
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Re: Isolating on agar from ms question [Re: Mushmix]
    #26355325 - 12/01/19 10:31 AM (4 years, 2 months ago)

You can do whatever you want lol. I would personally grow, clone and test clones, then keep the good one.


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OfflineMushmix
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Re: Isolating on agar from ms question [Re: bodhisatta]
    #26357418 - 12/02/19 03:26 PM (4 years, 2 months ago)

Just go right from ms to spawn jar then?


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InvisibleSmartattack
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Re: Isolating on agar from ms question [Re: Mushmix]
    #26357431 - 12/02/19 03:32 PM (4 years, 2 months ago)

That's not what he's saying at all lol. Use agar, don't sweat getting an isolated.


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OfflineHarperLewis
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Re: Isolating on agar from ms question [Re: Smartattack]
    #26357442 - 12/02/19 03:41 PM (4 years, 2 months ago)

Quote:

Mushmix said:
Just go right from ms to spawn jar then?




If it were me I'd go spore syringe to brf cakes. Either fruit them as is or spawn them to shoe boxes.

From there clone the fastest growing, largest, and or mushrooms that are producing nice clusters.

Transfer a couple of times  while trying to isolate for positive characteristics.

Then knock up some grain jars with your agar, and run a side by side trial of a couple tubs to see which performs the best.

Clone from there, and you should be able to use your newly isolated culture for storage and future grain inoculation / G2G transfers.


Edited by HarperLewis (12/02/19 03:43 PM)


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OfflineA.k.aM
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Re: Isolating on agar from ms question [Re: HarperLewis]
    #26357591 - 12/02/19 05:27 PM (4 years, 2 months ago)

You should read the penis envy isolation thread. It took him like 100 transfers and he had a handful of isolates and they all were garbage. Turns out variety is important. I think the current theory is that something like 8% of isolates will be worth keeping.

I’m kind of at the same point as you are I just started agar two days ago. I read A LOT and decided to go the clone route to start.

If you do go for an isolate the main advice from that thread is to dilute your spores to an extreme level first. Like put a drop from the syringe in a shot glass then fill it up and mix, then take a drop from there to agar, or possibly dilute that drop again.


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Edited by A.k.a (12/02/19 05:30 PM)


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Offlinetrippleblack
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Re: Isolating on agar from ms question [Re: HarperLewis]
    #26357600 - 12/02/19 05:32 PM (4 years, 2 months ago)

I always found the spore solution hit or miss on agar. if not fairly clean you'll get splotches of bacteria all over the plate. 

I would place one drop on a inoculation loop then dab the inoculation loop onto multiple petris of agar. transfer as soon as possible..

I usually get no more than 10 sectors when doing it like this.  usually a couple sectors of isolates out of 10 sectors will be keepers.  I usually grow out 15 or so good looking sectors..  I never liked cloning over isolating, when isolating sectors you have a lot more control over the outcome, the fruits are usually more unique and special too.


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