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Agar Isolate Q's (with pics)


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#1 abbr.

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Posted 20 March 2008 - 10:22 PM

I'm having a hard time seeing the sectoring.
Most of these pics are after 3 transfers I believe.
There was a few plates that I would look at,
and think it was all the same, then I look another
time and think it wasn't an isolated sub strain yet.
Any tips on how to tell?
Other than it all looking the same, lol.
You can see in the pics that I took wedges from them.
Figured I had the time, might as well go one more transfer.
Some other plates not pictured needed to be done anyway.
So, heres the pics from today, a few hours after transfers.

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#2 vinz

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Posted 20 March 2008 - 10:27 PM

those plates look good..
if you wanna go to substrain isolating, you have to transfer even smaller strands of myc
like grain size.. just get a strand of rhizo or something
then isolate into several plates
then keep on isolating
what strain is that?

#3 abbr.

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Posted 20 March 2008 - 10:37 PM

well, thats kind of what I've started doing.
Those are only a few of the plates.
So you're saying much smaller wedges?
Do you use an innoc loop, or cut with an exacto?
I've been using an exacto hobby knife.
So, what about isolates, am I anywhere near isolated yet?
I'm having a hard time deciding what's sectoring
and whats just rhizos clumping.
idk, I guess Ill just have to compare to the new transfers
and see what they look like then.

I'm not sure which plate is which strain,
like I said I have a bunch, but for the most part
they're Dixieland, Thai Pink, and Equador.
Those were just some random pics I took for reference here.

Thanks for takin' a look

#4 vinz

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Posted 20 March 2008 - 10:47 PM

i use the exacto knife
just get even smaller wedges, some dont even have to be wedges
you can cut out just the myc on top of the agar
and it will still grow

if you plan on isolating, you need to do tests
with all those plates, you can see which has the fastest and best rhizo growths
then test those agar plates into grain then spawn to subs
then take note of which plate you grew out and see if you like the results

more or less thats what you can do

also you can cut myc asap, like once you see even just an inch or so of growth then you can isolate already..

#5 abbr.

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Posted 21 March 2008 - 09:33 AM

So, what about isolates, am I anywhere near isolated yet?
I'm having a hard time deciding what's sectoring
and whats just rhizos clumping.



Bump for a little more insight

#6 warriorsoul

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Posted 21 March 2008 - 09:36 AM

You could always clone your best mushroom to get your isolate.

#7 abbr.

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Posted 21 March 2008 - 09:47 AM

I understand that, but thats not what were doing here.

#8 warriorsoul

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Posted 21 March 2008 - 09:55 AM

If your trying to get your best isolate..the agar method is inferior.
Is that what your doing here?

#9 fedshtkpndrk

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Posted 21 March 2008 - 06:06 PM

Your plates look great, you will need to take a small piece like vinz was saying to get transfers without sectoring. I recommend using a sterile needle to select a small piece of myc, sterile hobby knives work well too.

-feds

#10 CoyoteMesc

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Posted 21 March 2008 - 06:40 PM

yeah, like vinz and feds and freaky says...gotta get the pieces smaller. Looks like your taking too big of a piece and getting many sub-s.

Looks awesome btw :thumbup:

#11 onediadem

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Posted 21 March 2008 - 10:17 PM

yeah, like vinz and feds and freaky says...gotta get the pieces smaller. Looks like your taking too big of a piece and getting many sub-s.

Looks awesome btw :thumbup:


Agreed, very nice plates and what Feds said, a needle tip works great for pulling small sectors.. Whats your agar recipe abbr.? Those look nice and rhizy - nice work, keep us updated on your next transfer.

I don't see how agar is inferior Warrior. Clones can cut to the chase in some circumstances - If what you have in clone is already what you want.


#12 sooshane

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Posted 21 March 2008 - 10:42 PM

If your trying to get your best isolate..the agar method is inferior.
Is that what your doing here?


Agar is your friend. Somethings that you may consider agar use for:

1. Rhizomorphism—fast growing vegetative mycelium.
2. Purity of the strain—lack of cottony sectors.
3. Cleanliness of the myceiia—lack of associated competitor organisms (bacteria, molds and
mites).
4. Response time to primodia formation conditions.
5. Number of primordia formed.
6. Proportion of primordia formed that grow to maturity.
7. Size, shape and/or color of fruitbodies.
8. Total yield.
9. Disease resistance.
10. C02 tolerance/sensitivity.
11. Temperature limits.
12. Ease of harvesting.

Not my list but a good top twelve reasons to use agar.

I agree that a good clone can get you most of that but agar can help fine tune that which you want.

Nice work abbr.
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#13 warriorsoul

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Posted 22 March 2008 - 09:53 AM

I reason i like cloning better is because you have more control over the phenotype.
But i do like to work with agar for certain things,cleaning,storage...
Nice looking culture btw.:bow:

#14 abbr.

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Posted 22 March 2008 - 10:19 AM

If your trying to get your best isolate..the agar method is inferior.
Is that what your doing here?


Yes and no.
Its the whole process that I enjoy, its learning,
experimenting, a hobby!
My goal is to isolate several pure sub strains.
Then go through the fruiting process, and see
which fruit well.
I understand I could clone an impressive fruit,
and skip the whole isolating process, and I probably
will do that if a particular fruit, or cluster catches my eye.

What do you feel agar is inferior to? LC cloning?

OK, thanks for the responses everyone.
As you can see in the pics. I already transferred, so on
the next go around Ill go for much smaller strands.
I'll try to keep y'all updated with pics.

feds, I tried using a needle to clone from a fruit, like
you explained in your thread, I think I need a smaller gauge needle.
How do you use a needle tip for transfers?
Just kinda scrape up a lil piece?

"Whats your agar recipe abbr.?"

Pre mixed PDA

Wheat Malt Extract + Nut. Yeast

Light Malt Extract + Nut. Yeast

For the Light and Wheat I use...

750ml DH2O (My case of plates are in sleeves of 25, so I make 750ml and end up with a lil extra)
22.5g LME, or WME
15g Agar

And some have a gram of Nut. Yeast, some don't.

It seemed the yeast didn't break down in the mix,
when I ran it through the strainer before PC
it didn't filter out the left behind yeast.
Then when I poured the plates the yeast settled to
the bottom of the plates.
Is there any tips anyone has on the yeast?
Do I need to cook it longer?
Strain it with a cheesecloth maybe?
I haven't yet noticed a difference in the plates that
don't have it compared to those that do.

#15 warriorsoul

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Posted 22 March 2008 - 12:24 PM

Sorry, my post should have been clearer..
Its not the agar per say..
If i'm trying to get my best isolate i prefer to be able to see all the phenotypes and then choose.
By sectoring and isolating on agar the process is reversed, and you have less genetic variation to choose from.

A larger gene pool will increase your odds of finding a phenotype approaching the ideal.
:rasta:

#16 abbr.

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Posted 28 March 2008 - 09:16 PM

Heres some pics of Pan. Trop. and Goliath.
I thought it was strange how it seems to "eat"
the agar, anyone else notice this with Pans.?
Is it normal?
I thought in the archives I saw someone else
with a Pan.Cyan. plate that looked similar.

This first pic is the bottom of the Pan Trop....
 

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This is the top of that plate.....
 

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And this is the bottom of a Goliath plate.....

 

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The other pic is the top of a couple Goliath plates.
Thanks for looking!

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Edited by Sidestreet, 14 August 2015 - 10:29 AM.


#17 Myc

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Posted 29 March 2008 - 10:51 AM

Nice plates. Looks like you're having fun.
Is that a contam on your pan. trop. plate? The little blob off by itself. If so, you might want to get that outta there.

#18 abbr.

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Posted 29 March 2008 - 04:39 PM

yes, I believe that is contam, I was
just using that plate as reference to the
way the mycelium looks.
There was 2 Pan Trops to start,
the other one was used to nocc a few more,
those are the ones Ill use.
Im kinda just waiting to see what happens
when the mycelium reaches that contam.
The contam hasn't really grown, the mycelium
continues to plug along.

#19 Myc

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Posted 29 March 2008 - 08:41 PM

I'm discovering the wide world of contaminants myself. I've been using them as opportunities to experiment like yourself.

Very nice looking project. Never seen agar look like it's being eaten before.




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