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OfflineRotnpins
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Plate Pins- How to Clone & Test in One Easy Session * 17
    #28031270 - 11/03/22 02:48 PM (1 year, 2 months ago)

This write-up is a quick snapshot of how I clone and test my plate pins in one easy flow hood/SAB session.

As far as I know, the golden standard for plate pin procedure is to just transfer to a new agar plate, wait for it to revert back to mycelium and grow out on the plate, and then send a sample to your test jar(s) so you can test the clone culture and see how it performs..

As with everything else in this hobby, the process can be rather time consuming, especially if the culture doesn't perform to your liking.

I decided to combine the testing and cloning stages with a very simple proccess. Using this technique usually reduces the time it takes,  from when you transfer the plin to a new agar plate, to spawning your test tubs, by a week or more.

Supplies Needed

Agar plate with plate pin
New agar plate
Grain jar
Scalpel
Tweezers
Standard sterile work equipment (FFU/LFH/SAB, torch, 70% isopropyl alcohol)

Here is a picture of my work station before starting:



Here is a picture of the plate pin I have selected to use:


Note: This is easier with a medium/large sized plate pin. However, any size plate pin will work. Smaller pins will just take longer for the test jar to colonize. When starting with a smaller pin, I usually do an extra jar shake once I see new growth, that way there are more inoculation points to speed up the process.

The Technique (sterile technique will need slight modification when done in SAB)

Open the agar plate with plins, and the plate that you plan to transfer to:


Use the tweezers to remove plate pin. You may need to use a scalpel to remove any extra mycelium that is stuck to the plin.. You only want to transfer the pin. Any extra mycelium that you don't remove will potentially carry unwanted genetic diversity. So be sure and limit your transfer to the actual plin/clone tissue.



Move the plate pin to the new agar plate:


Use your scalpel to cut the plate pin in to two pieces..


I cut this plin in half, for the sake of simplicity.. Keep in mind, that it may be beneficial to cut a smaller piece to keep on the plate, and transfer the larger piece to the test jar so it will colonize a little faster.




I decided to keep the base of the stipe on the transfer plate and use the part with the cap to inoculate the jar... my logic behind this, is that if any of the spores from the cap germinate, I would rather the diversity show up in the test jar. That way the plate will be a more pure clone sample, and even if some of the spores result in a little genetic diversity in the test tub, I should still get a good indication of how the clone culture will perform.



Inoculate the jar using the plin:



Be sure and label the jar and plate in a way that is easily identifiable "Phobos Plin-1 11/03/2022"
That way, if the tub performs well, you can easily go back to the agar plate and expand the culture however you'd like.


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InvisibleShrimps
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Registered: 10/13/22
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Re: Plate Pins- How to Clone & Test in One Easy Session [Re: Rotnpins] * 2
    #28031281 - 11/03/22 02:54 PM (1 year, 2 months ago)

Awesome man! Right in time when I started wondering about this :laugh:

Love you man, great work! <3


--------------------
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Offline4-HO-Gumbo
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Re: Plate Pins- How to Clone & Test in One Easy Session [Re: Rotnpins] * 2
    #28031295 - 11/03/22 03:04 PM (1 year, 2 months ago)

Awesome, got primordia on some of my first plates, may have to try this out.


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InvisibleeLShaMukO
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Re: Plate Pins- How to Clone & Test in One Easy Session [Re: 4-HO-Gumbo]
    #28032973 - 11/04/22 05:19 PM (1 year, 2 months ago)

how much performance variation are you guys getting from pin clones from your plates ? i can only attest for speed on agar plates, that does vary from different pins from the same plate for some reason


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OnlineB Traven
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Re: Plate Pins- How to Clone & Test in One Easy Session [Re: eLShaMukO] * 1
    #28032984 - 11/04/22 05:24 PM (1 year, 2 months ago)

Filing this one under "why didn't I think of that?"

Sick shortcut!


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OfflineRotnpins
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Re: Plate Pins- How to Clone & Test in One Easy Session [Re: eLShaMukO]
    #28032985 - 11/04/22 05:25 PM (1 year, 2 months ago)

Quote:

eLShaMukO said:
how much performance variation are you guys getting from pin clones from your plates ? i can only attest for speed on agar plates, that does vary from different pins from the same plate for some reason




If I understand your question.. that's due to the variety of genetics on the plate.  The new plate and jar are from the same pin, so it's the same clone culture


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OfflineRotnpins
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Re: Plate Pins- How to Clone & Test in One Easy Session [Re: B Traven]
    #28033053 - 11/04/22 06:07 PM (1 year, 2 months ago)

Quote:

B Traven said:
Filing this one under "why didn't I think of that?"

Sick shortcut!




:lol: yeah.. Ive been doing it for a little while, and wondering why I had never seen someone else mention doing something similar. It really simplifies the whole plate pin process


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OfflinePutrema
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Registered: 02/20/20
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Re: Plate Pins- How to Clone & Test in One Easy Session [Re: Rotnpins]
    #28033290 - 11/04/22 08:13 PM (1 year, 2 months ago)

Hey, I came from the Ask Quick Questions thread where you have suggested this your new thread. Wow, it was unexpectedly inspiring. Are the photos taken currently? Can one see the progress of that very mycelium tissue in the near future? Have you some thoughts on throwing a pin directly into new media (agar, rye...) like you did here vs. removing some tissue from the inner mushroom and then putting it to new media? :sporedance:


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OfflineRotnpins
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Re: Plate Pins- How to Clone & Test in One Easy Session [Re: Putrema]
    #28033306 - 11/04/22 08:22 PM (1 year, 2 months ago)

Quote:

Putrema said:
Hey, I came from the Ask Quick Questions thread where you have suggested this your new thread. Wow, it was unexpectedly inspiring. Are the photos taken currently? Can one see the progress of that very mycelium tissue in the near future? Have you some thoughts on throwing a pin directly into new media (agar, rye...) like you did here vs. removing some tissue from the inner mushroom and then putting it to new media? :sporedance:




Glad you enjoyed the write up :super:

Yes.. those pictures were taken during my agar session yesterday. I'll be sure and post some update pictures in a few days.

As far as transferring pins to new media.. as long as they're plate pins, like the one I used, they should be sterile and result in a clean clone culture..

I have had plate pins pop up on contaminated plates.. in that case, I'll take a tissue sample so the contamination doesn't hitchhike to the new plate/jar.

With non plate pins, it's usually best to just take a tissue sample.. as long as your sterile technique is good, internal clone tissue should provide you with a clean plate without the need for transfers...

For some smaller species, I've been playing around with just throwing a pin or a cutting from a pin (not internal tissue) to a water agar plate.. this can simplify the process of cloning smaller fruits with delicate stipes.. you can read more about this in the water agar write up in my signature.


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OfflinePutrema
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Re: Plate Pins- How to Clone & Test in One Easy Session [Re: Rotnpins] * 2
    #28033353 - 11/04/22 08:49 PM (1 year, 2 months ago)

This makes sense. I currently have nothing activeley going on, except the fact, that there is some mycelium in the fridge and spores that are aging. Im looking forward the day, when I will let self collected spores thrive to mushrooms. But until then, I will see what I can do with living mycelium thats already there.

Yeah, I have read the first page of your water agar recently already. :smile:
Cause we now have a somewhat 'personal' connection, I will read the rest of the thread also I guess.


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InvisibleWinston Smith
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Registered: 09/03/22
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Re: Plate Pins- How to Clone & Test in One Easy Session [Re: Rotnpins] * 2
    #28033361 - 11/04/22 08:53 PM (1 year, 2 months ago)

Badass! I knew this was possible I just never had a jar ready when taking plins ha! Thanks for being the guinea, Rotnpins! You da cat!


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OfflineAugusto
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Re: Plate Pins- How to Clone & Test in One Easy Session [Re: Winston Smith] * 1
    #28034774 - 11/05/22 04:28 PM (1 year, 2 months ago)

Top dog man! I just have my first pins on plates (with average 10 primordia + 2 pins per plate, I will assume plates are clean), and was wondering what to do with em aparr from checking them out 'cause they're cool.

Will def prep some jars and try this out :rockon:


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OfflineRotnpins
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Re: Plate Pins- How to Clone & Test in One Easy Session [Re: Rotnpins]
    #28034792 - 11/05/22 04:36 PM (1 year, 2 months ago)

Once you guys give it a try, feel free to post fome pictures of your results in this thread.. it always makes things more interesting seeing everyone's different experiences and results using a particular technique


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InvisibleeLShaMukO
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Re: Plate Pins- How to Clone & Test in One Easy Session [Re: Rotnpins]
    #28034869 - 11/05/22 05:09 PM (1 year, 2 months ago)

Quote:

Rotnpins said:
Quote:

eLShaMukO said:
how much performance variation are you guys getting from pin clones from your plates ? i can only attest for speed on agar plates, that does vary from different pins from the same plate for some reason




If I understand your question.. that's due to the variety of genetics on the plate.  The new plate and jar are from the same pin, so it's the same clone culture




thanks for responding, should have specified on my behave on how much variation are people getting from taking Pins from MS plates as i have read posts also about cloning pins from an agar plate is not cloning fruits rather just transfers?

just wondering since id assume the difference on speed im seeing is cause of genetic variation like you mentioned before.


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OfflineSirPsycho
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Re: Plate Pins- How to Clone & Test in One Easy Session [Re: Rotnpins]
    #28034935 - 11/05/22 05:45 PM (1 year, 2 months ago)

Quote:

Rotnpins said:
and wondering why I had never seen someone else mention doing something similar



Best practice and all that. Same reason you should test your LC on agar every time.

Like I said in the other thread though, it's common practice among certain gourmet cultivators. Faht also chimed in and is something he does.

I like the spirit, I'm totally with stretching the limits of best practices.


--------------------
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OfflineRotnpins
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Re: Plate Pins- How to Clone & Test in One Easy Session [Re: SirPsycho]
    #28034984 - 11/05/22 06:05 PM (1 year, 2 months ago)

Quote:

SirPsycho said:
Quote:

Rotnpins said:
and wondering why I had never seen someone else mention doing something similar



Best practice and all that. Same reason you should test your LC on agar every time.

Like I said in the other thread though, it's common practice among certain gourmet cultivators. Faht also chimed in and is something he does.

I like the spirit, I'm totally with stretching the limits of best practices.




I get where you're coming from... but, at the same time, I don't see a point in testing it..

As long as the donor plates that the pins grew on were clean, and your sterile technique is half way decent, I don't see where there would be any more opportunity for contam than if you were transferring a wedge from a clean plate..

If I was doing tissue samples from fruits and sending them straight to grain, I could see
it not being best practice.. there is always a small chance of bacteria.. but plate pins are being transferred from a clean environment to grain.


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OfflineAugusto
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Re: Plate Pins- How to Clone & Test in One Easy Session [Re: Rotnpins]
    #28035628 - 11/05/22 10:12 PM (1 year, 2 months ago)

These are my boys. I can't count for shit, there are at least 30+ knots per plate. Gonna let these boys grow for a couple of days, then to grain and maybe agar (although I really do hate agar work in my SAB). Will post back here :thumbup::mushroom2:


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OfflineSirPsycho
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Re: Plate Pins- How to Clone & Test in One Easy Session [Re: Rotnpins]
    #28035950 - 11/06/22 04:05 AM (1 year, 2 months ago)

Quote:

Rotnpins said:

As long as the donor plates that the pins grew on were clean, and your sterile technique is half way decent, I don't see where there would be any more opportunity for contam than if you were transferring a wedge from a clean plate..





There isn't any more opportunity or any difference.

Quote:

Rotnpins said:
If I was doing tissue samples from fruits and sending them straight to grain, I could see
it not being best practice.. there is always a small chance of bacteria.. but plate pins are being transferred from a clean environment to grain.



Even with a regular fruit, you'd still be ok if your sterile technique is good.


--------------------
:pm:Ask me about free Ps tampanesis, Ps subtropicalis and Ps cubensis (ESS) prints:pm:
Balance in life is like running on ice.

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OfflinePhilberto
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Re: Plate Pins- How to Clone & Test in One Easy Session [Re: SirPsycho]
    #28036035 - 11/06/22 06:15 AM (1 year, 2 months ago)

Not sure I understand why pins are coming up. This is just mycelium growing across a plate
Right?

I haven't got very far, but I do have some mycelium growing on plates just to see if it's clean. Can I expect pins just as a matter of course?


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OfflineSirPsycho
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Re: Plate Pins- How to Clone & Test in One Easy Session [Re: Philberto]
    #28036041 - 11/06/22 06:22 AM (1 year, 2 months ago)

Quote:

Philberto said:
Not sure I understand why pins are coming up. This is just mycelium growing across a plate
Right?

I haven't got very far, but I do have some mycelium growing on plates just to see if it's clean. Can I expect pins just as a matter of course?



Fungi will attempt to fruit and reproduce whenever they have run out of nutrients. This will happen in pretty much any given situation. They'll fruit inside spawn jars that sit too long, they'll fruit on agar plates, they'll even fruit in liquid culture.


--------------------
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Balance in life is like running on ice.

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