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Catattack Some Agar (again)


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#81 Heirloom

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Posted 11 January 2016 - 12:33 PM

I have been using my pc for several days preparing agar, to clean a few and to get more pure cultures.
 I learn from these posts and other on agar, they helped me to try agar and get as far as I have.

Cat do you expect to get fruits from your all your PE clones or only a few? Thanks I just want to learn. good luck
 


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#82 CatsAndBats

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Posted 11 January 2016 - 12:43 PM

I have been using my pc for several days preparing agar, to clean a few and to get more pure cultures.
 I learn from these posts and other on agar, they helped me to try agar and get as far as I have.

Cat do you expect to get fruits from your all your PE clones or only a few? Thanks I just want to learn. good luck
 

Will you please rephrase that? Do I expect all of my jars to be clean and utilized for grows, no. Did I answer right?


Edited by catattack, 11 January 2016 - 12:46 PM.

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#83 Heirloom

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Posted 11 January 2016 - 02:59 PM

cat I cloned PE and my contaminated jars were destroyed.  I did not expect you have 100% pure agar work, no one does.
I never have.


I only mean that when you transfer a pure culture to grain an grow it out on grain do they all produce? I hope that's clear.

I cloned some PE  just transferred some wedges to grain , if the jars grow clean can I expect them to fruit?

 I was just asking not in any way being negative, I am here to learn.

I was thinking clones and isolates might be similar, obviously not.

accept my apology for any misunderstanding. I hope this is clear



 


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#84 CatsAndBats

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Posted 11 January 2016 - 03:11 PM

Dude, it's me. Stop apologizing!  I learned that for it to be a true isolate, it'll have perfectly radial growth and merge seamlessly with another version of itself. taking a piece and putting it to agar cleanly I think technically is still a 'clone' ie 9er clone tek.

 

It should fruit IMO/E, but PE is a cruel mistress, so...

 

@peace @mlb @coorsmikey @whitethumb (or anyone else who knows definitively) will you please double check the above? 


Edited by catattack, 11 January 2016 - 03:12 PM.

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#85 Heirloom

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Posted 11 January 2016 - 03:35 PM

Thanks cat .

Heres a pic of an agar jar with a clone of pe growing for a while.  when I cut the agar and put into jars I can expect to get something?. The  spot on the end of the pic is the small pe tissue.

the other agar jar is cut up and who wants to see that.

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  • PE 111 002.JPG

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#86 CatsAndBats

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Posted 11 January 2016 - 03:42 PM

Thanks cat .

Heres a pic of an agar jar with a clone of pe growing for a while.  when I cut the agar and put into jars I can expect to get something?. The  spot on the end of the pic is the small pe tissue.

the other agar jar is cut up and who wants to see that.

Here's one of my agar wedges (PE clone)

DSC02020.jpg

 

Here's some good rhizo off of said clone:

DSC02022.jpg


Edited by catattack, 11 January 2016 - 03:56 PM.

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#87 CatsAndBats

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Posted 15 January 2016 - 06:41 PM

Hey y'all. I need some more eyes on these please! These are the PE clones off of a big ol cluster, off of the biggest fruit. How should I proceed?

 

Each photo is it's own jar. Should I chose one set of rhizo and try to grow that out in multiples? Or keep 'training' it somehow?

 

DSC02028.JPG

 

 

 

DSC02029.JPG

 

 

DSC02030.JPG


Edited by catattack, 15 January 2016 - 06:43 PM.

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#88 CatsAndBats

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Posted 16 January 2016 - 11:43 AM

ATTENTION AGAR HEADS, please get some eyes on the last post, any suggestions will be appreciated.

 

I'm looking at y'all, @peacefrog (where does that profile name come from?) and @whitethumb.

 

;)


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#89 meyer

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Posted 16 January 2016 - 03:49 PM

I wish I could help you cat, funny I just asked. A few people about agar.

#90 MLBjammer

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Posted 16 January 2016 - 05:40 PM

If you are looking for a true isolate, you will want to sector it and transfer some more.  Tough to tell from pics sometimes, but it looks like more than one culture in each of the above jars.

 

Sorry for the late response.  Busy Saturday.


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#91 CatsAndBats

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Posted 16 January 2016 - 05:50 PM

If you are looking for a true isolate, you will want to sector it and transfer some more.  Tough to tell from pics sometimes, but it looks like more than one culture in each of the above jars.

 

Sorry for the late response.  Busy Saturday.

 

So grab from one sector and make say 5 jars and do it again and again until it's a perfect circle right? Do I try to "marry" them later and see if they join seamlessly?


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#92 sgfcchamber

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Posted 16 January 2016 - 05:55 PM

Hey y'all. I need some more eyes on these please! These are the PE clones off of a big ol cluster, off of the biggest fruit. How should I proceed?

 

Each photo is it's own jar. Should I chose one set of rhizo and try to grow that out in multiples? Or keep 'training' it somehow?

 

I see sectors in each of those photos, so if it were me, I'd continue with additional transfers until you have each sector in each jar as it's own monoculture and fruit out each monoculture individually.  Not necessarily an entire tub for each one though.  When I was isolating strains within clones what I would do would be what I just described, and then I'd use a wedge from each of the isolates to innoculate a PINT jar of grain (the same grain I'd be using if/when I got around to using that isolate in 'en masse).

 

Then I'd take each colonized PINT jar of grain (each having it's own isolate/monoculture) and make "big cakes" in a 2 quart tupperware container (2 quarts = 4 pints).  So the cakes would be 3 parts bulk substrate and then the pint of grain.  The substrate should be the same "recipe" that you'd use should you choose to take that monoculture/isolate to a true bulk (monotub).  Reason being is that what results you get from the mini-cake could be substrate dependent (to some degree), so if you change the recipe when you go bulk you could end up with entirely/slightly different results because it grew well/poorly/mediocre on one substrate but behaves differently on another.

 

I'd then take these cakes and fruit them, simultaneously, in the same fruiting chamber (to ensure that the conditions are the same for each cake and that your results aren't skewed by environmental variables by having them in separate containers/fruiting the at different times).

 

Take notes on everything, such as time to pin, total yield, number of tight clusters, pins per sq in (if you're inclined for that level of detail) and so on.

 

The above is from experience and a lot of trial and error cloning & isolating.


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#93 CatsAndBats

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Posted 16 January 2016 - 06:16 PM

That is a meaty answer, I love it, thank you!

 

A little trick I use, I include a small amount of everything that my mycelium might encounter through it's entire life cycle. I have a couple tupperwares that have my only true supplement, which is powdered egg shells (calcium and a little nitrogen boost). To that I add rice, coir, old trim from former grows, oats etc, powder the shit out of it in a coffee grinder, and everything gets anywhere from a pinch (PF/BRF cake) to handfuls (large mono). The reasoning behind this is the mycelium build the necessary enzymes in advance to ease their transition to a different sub.

 

 

Thanks again @sgfc!


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#94 sgfcchamber

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Posted 16 January 2016 - 07:04 PM

That is a meaty answer, I love it, thank you!

 

A little trick I use, I include a small amount of everything that my mycelium might encounter through it's entire life cycle. I have a couple tupperwares that have my only true supplement, which is powdered egg shells (calcium and a little nitrogen boost). To that I add rice, coir, old trim from former grows, oats etc, powder the shit out of it in a coffee grinder, and everything gets anywhere from a pinch (PF/BRF cake) to handfuls (large mono). The reasoning behind this is the mycelium build the necessary enzymes in advance to ease their transition to a different sub.

 

 

Thanks again @sgfc!

 

That is a very clever idea...I like it, a lot!  I believe I'll begin doing that with my agar :)

 

As for isolate testing and finding a "winner" have a look at this (run through it using only 2 isolates to get a feel for how it works):  http://substratecalc...om/isolate.html


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#95 peacefrog

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Posted 16 January 2016 - 10:52 PM

ATTENTION AGAR HEADS, please get some eyes on the last post, any suggestions will be appreciated.
 
I'm looking at y'all, @peacefrog (where does that profile name come from?) and @whitethumb.
 
;)


Like said above, more isolating is needed there, cat. But good looking clone! Keep us updated on your progress.

And the name comes from one of my favorite Doors tunes.
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#96 MLBjammer

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Posted 17 January 2016 - 06:38 AM

"Peace Frog" The Doors

 

[Direct Link]


Edited by MLBjammer, 17 January 2016 - 06:39 AM.

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#97 peacefrog

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Posted 17 January 2016 - 06:41 AM

That's it, jammer! Love that song.
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#98 Microbe

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Posted 17 January 2016 - 12:23 PM

uploadfromtaptalk1453050601584.jpg uploadfromtaptalk1453050606608.jpg uploadfromtaptalk1453050612063.jpg

Dont have much time and its really hard to tell by the pics. If its not to late, i would select from the arrows in your pictures above if it were me.

The problem with clones and even transferring on agar is that you really need a scope to look for sectoring. Allthough it is easy to assume we established a mono-culture, it is possible that sectoring exists but we cannot see it clearly as we would from a spore germination.

However and for our purposes, if you have a plate that you cant clearly see any sectoring, and the culture has proven to fruit, then i would declare it a mono and would slant it. It is very possible to obtain a true isolate after a single transfer of a very small selection grown out from a clone.

When i select i look for 1st speed then followed by fruiting. So if those were my clones i would make sure that they were placed to the the center of the jar then you can easily see the fastest growth. Other factors come into play and could throw it out of whack such as inconsistent moisture content on the agar surface or nutrient pockets exist are a few.

Overall those clones look good.
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#99 MLBjammer

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Posted 17 January 2016 - 08:56 PM

Good observations, Microbe.

 

Here's my namesake:

 

[Direct Link]


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#100 coorsmikey

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Posted 17 January 2016 - 09:05 PM

Good observations, Microbe.
 
Here's my namesake:
 

[Direct Link]

Holy Crap Jammer! How the heck did I ever miss that connection? All this time I thought you were a baseball fan. :face palm:
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