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Cloning Pins From Agar to Agar
    #14731236 -

So i have this petri dish of Koh Samui Mycelium. It was inoculated about 5 weeks ago with a drop of spore solution.





Now it has all of these pins on it. I am about to clone a few of them to low-nute MEA and see what happens. I imagine it will be just like cloning a pin from a tub or tray.

I wondered if anyone had tried this before, cloning a pin from agar to agar, and what your results were.

Update 7/19



On 7/13 i pulled 3 pins off of this dish and places them individually on dishes of low-nutrient MEA.



I may do some transfers from here. Not really a big deal, it just crossed my mind. Plus mini mushrooms look realy cool.


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~You're already dead, but you're about to die~


Penis Envy in Monotubs      Ecuador in Monotubs

Edited by GungGung (07/19/11 02:03 PM)

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Re: Cloning Pins From Agar to Agar [Re: GungGung]
    #14792161 -

Updated 7/19


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~You're already dead, but you're about to die~


Penis Envy in Monotubs      Ecuador in Monotubs

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Re: Cloning Pins From Agar to Agar [Re: GungGung]
    #14792179 -

That is pretty cool.  I bet you get very prolific fruiting cultures.


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Of course, then there's the problem of eating vermiculite. On the bright side, it makes your poop and teeth glitter.  Just pretend it's christmas.
-RR

Those mushrooms are fine.  Your friend is a pussy.
-RR

Outdoor Patch

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Re: Cloning Pins From Agar to Agar [Re: M11]
    #14792290 -

No way to tell until i fruit them. I have KS multispore inoculated spawn near full colonization which i will be doing a grow with, and getting more clones from, but i will be testing these isolates as well.


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~You're already dead, but you're about to die~


Penis Envy in Monotubs      Ecuador in Monotubs

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Re: Cloning Pins From Agar to Agar [Re: GungGung]
    #14792362 -

I'll be doing the same thing this week. An easy way to make an isolation! Well done, and keep us posted on how she fruits when spawned to bulk, of you don't mind.


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Re: Cloning Pins From Agar to Agar [Re: Puma]
    #14792420 -

the strains that end up fruiting on your agar dish are said to be

"co2 tolerant" strains.


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AMU - AMU Q & A - MyVideo Teks!

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Re: Cloning Pins From Agar to Agar [Re: k00laid]
    #14792515 -

Thats cool. Since CO2 is vital for colonization, and the removal of it is the primary pinning trigger for cubensis, i figure that they are all pretty CO2 tolerant. Even CO2 dependant.

If i was planning on fruiting invitro, say bags or something, this would be helpful to me. I prefer to fruit in a monotub, which provides a huge ammount of fresh air to the developing fruits, after they have colonized the sub in the high CO2 environment of the taped up tub with no airflow.

Can you think of any distinct advantages of a substrain that is "CO2 tolerant"?


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~You're already dead, but you're about to die~


Penis Envy in Monotubs      Ecuador in Monotubs

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Re: Cloning Pins From Agar to Agar [Re: GungGung]
    #14792641 -

Cubensis is CO2 tolerant, period.

Transferring pins from one petri dish to another is redundant.  It also isn't cloning, but just transferring.  Use that dish to inoculate grains if you want to test the strain.
RR


--------------------
Download Let's Grow Mushrooms



semper in excretia sumus solim profundum variat

"I've never had a failed experiment.  I've only discovered 10,000 methods which do not work."
Thomas Edison

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Re: Cloning Pins From Agar to Agar [Re: k00laid]
    #14792675 -

Ok. So with all that being said, I'm assuming that (steril precession taken as much as possible) you could pull a pin off a BRF cake from a FC, and then place it on agar as a clone? Not that that is ideal because it could be an abort or some other issue. But for a fast ISO? Would you need to cut the pin up? Or could you just place it on the agar and expect results?


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I am a Noob. Can't help it. Try and forgive me for any NewB mistakes or comments. Lol and if you would like to mentor me, PM me please.

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Re: Cloning Pins From Agar to Agar [Re: hermandamerman]
    #14792687 -

hermandamerman said:
Ok. So with all that being said, I'm assuming that (steril precession taken as much as possible) you could pull a pin off a BRF cake from a FC, and then place it on agar as a clone? Not that that is ideal because it could be an abort or some other issue. But for a fast ISO? Would you need to cut the pin up? Or could you just place it on the agar and expect results?


Yea you can do this and it could already be an iso.  Clones usually have between 1-7 sectors.  Just place it on agar and transfer the sectors


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Pictures say 1,000 words.  What do these pics tell you?
Any mycolgy question answered by AMU.  Fast and good advice
Read all these teks and you can be elite.
Best teks out there!!!!!

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Re: Cloning Pins From Agar to Agar [Re: UnnamedGrower]
    #14793385 -

My whole point with this thread was to explore the concept of arriving at an isolate, without the presumably necessary step of colonizing grain and fruiting the multi-substrain mycelium and then cloning from those fruits, or having to do a ton of transfers.

When i did my Penis Envy isolation run on agar it took me an average of 10 transfers along each path to arrive at my isolates. That is a lot of petris and a lot of antibiotic MEA, energy to sterilize, etc... I am always striving to reduce waste.

My thought was that if one simply waited for a petri dish to fully colonize and pin, those pins would facilitate the production of isolates in the same way as an inner stem tissue clone of a mature mushroom or pin from a casing, which usually takes me 2 transfers to get to an isolate.

These pins are still individual mushrooms, just very small, and, since the dish was inoculated with a multispore solution, i presume that i could clone in the same way that i usually clone from tissue, by placing the tissue on agar and isolating until no sectoring is apparent.

I have no intention of using this technique to replace my current method of cloning, usually from a big pin in the center of a big cluster of healthy pins. I think observing a mushroom in its fruiting environment gives you the best idea of what traits you are actually cloning. Cloning these pins that grew on a petri dish, i have no way of knowing how big they would get, frequency of aborts, average flush size, and other variables. However, i only used 5 petris total on Koh Samui, and only handled it in my glovebox a few times. If it only takes me 2 more transfers to get to an isolate, like i think it will, then i have significantly reduced the amount of waste attributed to my hobby, something that i think is extremely important.

I already used another multispore petri to inoculate grain, and i will select other fruits to clone when i spawn that grain to bulk and fruit it.
I started a thread in Logs&Pics following my progress with this Koh Samui.


--------------------
~You're already dead, but you're about to die~


Penis Envy in Monotubs      Ecuador in Monotubs

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Re: Cloning Pins From Agar to Agar [Re: GungGung]
    #14794787 -

GungGung said:
Thats cool. Since CO2 is vital for colonization, and the removal of it is the primary pinning trigger for cubensis, i figure that they are all pretty CO2 tolerant.


har har.

i was referring to their ability to pin without the removal of co2 (like in your agar dishes) :omgz:


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AMU - AMU Q & A - MyVideo Teks!

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Re: Cloning Pins From Agar to Agar [Re: GungGung]
    #14794796 -

GungGung said:
Can you think of any distinct advantages of a substrain that is "CO2 tolerant"?


growing in bags.


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AMU - AMU Q & A - MyVideo Teks!

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Re: Cloning Pins From Agar to Agar [Re: k00laid]
    #14794993 -

Word, that is the first thing my mind went to. I have not yet done any bags, and do not think i will, i aspire to reduce waste, so adding non-decomposable autoclavable bags would be counterproductive for me.
I wonder if you started doing that on agar, then took the same isolate to grains/bulk and deprived them of fresh air while fruiting, took spore prints from that grow, did the same thing on agar again,re-isolated took it back to grains/bulk, maybe 3 or 4 more times, if you would breed in a CO2 tolerant trait, then fruit them regularly and see how they react to having fresh air after a few generations of deprivation. I wonder if it would have an effect on pin sets.
I have never considered breeding genetic traits into fungus through deprivation before, i know it works great with plants though. It makes sense, when i clone on agar i use low nutrient content agar and it speeds up growth drastically as the mycelium chases down the nutrients. I can only imagine that applying other stresses would cause other traits to surface. Has anyone tried this kind of thing?


--------------------
~You're already dead, but you're about to die~


Penis Envy in Monotubs      Ecuador in Monotubs

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Re: Cloning Pins From Agar to Agar [Re: GungGung]
    #14795116 -

GungGung said:
When i did my run on agar it took me an average of 10 transfers along each path to arrive at my isolates. That is a lot of petris and a lot of antibiotic MEA, energy to sterilize, etc... I am always striving to reduce waste.



Avoid antibiotic agar unless you have an active bacteria infection you're trying to isolate from.  Don't use it as standard procedure.
RR


--------------------
Download Let's Grow Mushrooms



semper in excretia sumus solim profundum variat

"I've never had a failed experiment.  I've only discovered 10,000 methods which do not work."
Thomas Edison

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Re: Cloning Pins From Agar to Agar [Re: RogerRabbit]
    #14795414 -

RogerRabbit said:
Transferring pins from one petri dish to another is redundant.  It also isn't cloning, but just transferring.  Use that dish to inoculate grains if you want to test the strain.
RR


I think I'm missing something here, RR. If taking a tissue sample from a pin or mushroom fruiting from a MS substrate is a way to isolate a clone, then why wouldn't it be the same when taking a tissue sample from a pin or mushroom forming on MS petri dish? Isn't a pin a pin, and a mushroom a mushroom, and thus clone-able, no matter where the fruit forms?


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Re: Cloning Pins From Agar to Agar [Re: GungGung]
    #14795442 -

GungGung said:
Word, that is the first thing my mind went to. I have not yet done any bags, and do not think i will, i aspire to reduce waste, so adding non-decomposable autoclavable bags would be counterproductive for me.
I wonder if you started doing that on agar, then took the same isolate to grains/bulk and deprived them of fresh air while fruiting, took spore prints from that grow, did the same thing on agar again,re-isolated took it back to grains/bulk, maybe 3 or 4 more times, if you would breed in a CO2 tolerant trait, then fruit them regularly and see how they react to having fresh air after a few generations of deprivation. I wonder if it would have an effect on pin sets.
I have never considered breeding genetic traits into fungus through deprivation before, i know it works great with plants though. It makes sense, when i clone on agar i use low nutrient content agar and it speeds up growth drastically as the mycelium chases down the nutrients. I can only imagine that applying other stresses would cause other traits to surface. Has anyone tried this kind of thing?


Each time you grow anything from spores you have started from scratch. It's like writing an essay and getting half of what you want to say on paper and then ripping it up and starting over.

The only way to isolate the ideal qualities of what you are looking for is by testing each and growing it and seeing.


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My Old Grow Logs


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Re: Cloning Pins From Agar to Agar [Re: GungGung]
    #26420296 -

Hello gunggung, Very interesting post on PE isolation on agar and cloning pins from agar. I’m trying to resurrect a degrading PE culture but haven’t figured out the best approach. How were your results from taking a pin from a fruiting agar dish? Also would fruiting a multispore isolate right in the agar dish be a good way to bypass a lot of steps. I would imagine if it fruits well on agar it would fruit well on a nutritious substrate. I dunno, I’ve been proven wrong a lot on this long road to mushroom growing lol.

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Re: Cloning Pins From Agar to Agar [Re: mirage1910] * 1
    #26420298 -

idk if you noticed but this post is 8 years old

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Re: Cloning Pins From Agar to Agar *DELETED* [Re: RogerRabbit]
    #28353049 -

Post deleted by lolowiggs

Reason for deletion: Realized it was a silly question, dont want to clog an old thread


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Call me professor, I graduated with my M.S in BS as well as a dual major in recreational substance ingestion.

Lolo’s the name psychedelics are my game.

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