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Clone tissue to agar, not sure what do next


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#1 adrian118

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Posted 30 March 2021 - 11:12 PM

Burma grow fruited really nice and so I took some tissue from one and put them to agar. ( pics below ), not sure what to do now.

Thought was I'd make transfers and isolate, then use agar wedges to inoculate grain. But maybe these look good to go?

Or should I do a LC with one and keep the others in the fridge?

First time going this route, so many teks to choose from, wondering what you guys think or suggest.

 

Thank you all!

 

3/23

IMG_3181.jpg IMG_3182.jpg IMG_3183.jpg

 

3/30

IMG_3203.jpg IMG_3204.jpg IMG_3205.jpg


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#2 Auhron

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Posted 31 March 2021 - 12:01 AM

Why not try a bit of each? Only takes a syringe tips worth to start a Liquid Culture, you could start multiple and still use that plate to play with transfers to agar, use another plate to grain, and still have one for the fridge! 


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#3 Sidestreet

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Posted 31 March 2021 - 03:33 AM

Those look nice!  I like the little 2 oz portion cups as plates.  Could you tell us a little about how you're making them?  Are you using anything to seal/protect them, or are they only protected by the lids?

 

I also like the idea of putting at least one of them into cold storage, to make sure you have a backup of your clone.

 

I would fruit your clone, too, so you can see how it performs as-is.  If it is really good, you could take your backup and make several more backups to store so you are preserving the genetics.

 

Or you could fruit it and then take a spore print from some of the most desirable fruits with an eye toward repeating the process and further refining your strain.


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#4 rockyfungus

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Posted 31 March 2021 - 12:14 PM

I've had such hit or miss luck with those cups. Sometimes the lids will be fine other times the lids are not rated for PCing. When they work as no-pour, they rock!


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#5 adrian118

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Posted 31 March 2021 - 03:16 PM

Prepping some grain and LC today. Think I have some LME somewhere.

 

Hey Sidestreet,

These were done using PDA heated to a simmer on the stove then poured into a media bottle which was then PCd. Then the cups are poured and left to cool in a SAB. Nothing sealing the cups except for the lids which are very tight fitting. Trusting them to be sterile out of the bag since they are food grade. 

 

Thanks for the advise everyone! 


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#6 Arathu

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Posted 31 March 2021 - 04:52 PM

Those look like very happy clones. Nice work!

 

I would still make transfers from at least one of those plates and probably make transfers from a selection of those just to play things safe......

 

My process (regardless of how things made it to the plates in the first place) is definitely multiple transfers on agar and then agar wedges to sterile grains......G1

 

The most vigorous of those G1 jars is used as a grain master for G2G expansion, G2 and beyond, in preparation for spawning to the fruiting substrate....

 

So pretty much textbook Stamets.....which makes sense because I still keep the books handy for reference. Many times it's not even close to that extensive.

 

Some times I just feel that a fungus on the plate will grow and do it, put it to grains, it's a gamble (you win and loose when doing that). It is wise to be rigorous if making anything in bulk IMHO.    

 

Excellent stuff right here!

 

A


Edited by Arathu, 31 March 2021 - 05:13 PM.

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#7 Oldpunk

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Posted 31 March 2021 - 07:35 PM

Those look awesome! Im getting ready to try my first clone/agar attempt and can only hope they look that good.

Nice work!

Edited by Oldpunk, 31 March 2021 - 07:36 PM.


#8 FunnyFarmer

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Posted 01 April 2021 - 12:12 AM

I've been reading RR's notes and currently going through agar, cloning etc. A lot to wade though there especially with the redundant notes... Been musing whether or not to condense them into a more readable format, a Cliffs Notes edition if you will. A rather daunting task to keep from loosing important info. Anyway he would take a sample from the fastest mycelia growing from the center of inoculation and put that to another agar plate. He also concentrated on ropy growth as opposed to cottony growth taking samples from the leading edge of the fastest growth to nock another agar plate., etc, etc,etc. When he was satisfied he would then put a wedge to grain and let colonize putting the rest in the frldge for either more agar rounds or nocking up more grain if he liked the results.. This guy was a regular Dr. Demento when it came to raising shrooms.


Edited by FunnyFarmer, 01 April 2021 - 12:17 AM.

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#9 Sidestreet

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Posted 01 April 2021 - 04:21 AM

I like the idea of that project, FF.  "Abridged Rabbit (Compiled and Edited by FF)". 

 

It's hard for people to know where the good stuff is--where the good threads and information are--without being around forever and spending a ton of time digging.  Kudos to those who have done it, but I think it's a worthy pursuit to gather all that stuff together for others' benefit.  It seems good for the OMC at-large.


Edited by Sidestreet, 01 April 2021 - 04:26 AM.

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#10 adrian118

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Posted 07 April 2021 - 03:52 PM

Made a couple transfers.

 

April 1

IMG_3213.jpg IMG_3216.jpg IMG_3217.jpg

 

Today

IMG_3238.jpg IMG_3239.jpg

 


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#11 Arathu

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Posted 07 April 2021 - 05:54 PM

Indeed..... :thumbs_up:

 

Those look good from here.....

 

A



#12 FunnyFarmer

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Posted 08 April 2021 - 10:56 AM

I decided to make an attempt at consolidating RR's notes but concerned about possible conflicts with the Shroomery forum. Thoughts anyone? Don't get your hopes up though, I said attempt and I'm not a writer and my style is really dry, to the point. This is a rather daunting task considering that not everything is in one area, a lot is but... I guess I'm afraid that I'll miss something important, the purists will do their thing, , , Wiley said it best with this comic strip from Non Sequitur
NonSequiter_Grotto.gif

I use it in my signature on a motorcycle forum. Hmmm...

Edited by FunnyFarmer, 08 April 2021 - 10:57 AM.





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