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Offlinemetaphoric
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Tissue cloning question
    #26380092 - 12/13/19 08:03 PM (4 years, 1 month ago)

Well I'm finally getting into agar. I've always just done g2g with ms masters. I want to clone a large fruit clump. I've done a lot of searching and read plenty of teks but every tek pretty much skips over the tissue extraction/transfer step. This gives me hope since it must be as simple as it seems but my question is should I toss the mushroom in alcohol or hydrogen peroxide before cutting into it and extracting a piece of flesh from the middle?


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:mushroom::mushroom::mushroom::mushroom::mushroom::mushroom::mushroom::mushroom::mushroom::mushroom:
:mushroom:                  :mushroom2:                :mushroom:
:mushroom:            :mushroom2::mushroom2::mushroom2:            :mushroom:
:mushroom:      :mushroom2::mushroom2::mushroom2::mushroom2::mushroom2:      :mushroom:
:mushroom::cookiemonster: BEHOLD :elmo::mushroom:
:mushroom::mushroom::mushroom::mushroom::mushroom::mushroom::mushroom::mushroom::mushroom::mushroom:


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Offlinesan pedro guy
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Re: Tissue cloning question [Re: metaphoric]
    #26380093 - 12/13/19 08:06 PM (4 years, 1 month ago)

No.

Just make sure you get a clean piece from the center of the stipe.

If you have smaller ones you don’t care about it couldn’t hurt to practice on them before you try the one you wish to clone.

Gluck mate!


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Noob Grow Along 2022


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OfflinecronicrFacebook
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Re: Tissue cloning question [Re: san pedro guy] * 1
    #26380096 - 12/13/19 08:10 PM (4 years, 1 month ago)

Rip the fruit open don't cut it open and take a piece of flesh


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It doesn't matter what i think of you...all that matters is clean spawn

I'm tired do me a favor


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Offlinedaysbetween
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Re: Tissue cloning question [Re: cronicr]
    #26380099 - 12/13/19 08:11 PM (4 years, 1 month ago)

rip it open, sterilize your blade, and cut out a small chunk from the inside.


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InvisibleStygianKnight
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Re: Tissue cloning question [Re: san pedro guy] * 1
    #26380109 - 12/13/19 08:16 PM (4 years, 1 month ago)

No, just pick a healthy looking fruit.

Squeeze the stipe so it splits and you can peel the fruit in half.  The center stipe material is very stringy, make two horizontal cuts to sever the strings and then peel up the chunk.  The mycelia can be sticky so focusing on clean cuts can make picking it up easier.  Practicing on a few fruits is a good idea so you can understand how it might try and fight you.  When placing on agar it can sometimes be easier to cut into the agar and drag the blade to deposit the tissue.  Expect to transfer to a new plate once it’s grown out a bit.


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OfflineJohnRainy
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Re: Tissue cloning question [Re: metaphoric]
    #26380249 - 12/13/19 09:57 PM (4 years, 1 month ago)

I find the caps are the most convenient place to take from.  I put a little slit on the bottom of the stem first and then tear it right in half up from there.  The cap's flesh just a little above the gills in the centre has a lot of room for your scalpel and it's deep flesh so you won't poke out the other side like can happen on the stem.

The flesh there also seems firmer to me and less likely to die from bruising too.  Sometimes a sample will just turn blue and be dead on the agar and it happens more with stem samples IME.

I just take two samples per mushroom, one from each half of the cap.

You'll want to do this before the cap begins to open, or you might get spores mixed in with your work and you don't want that.


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Offlinemetaphoric
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Re: Tissue cloning question [Re: StygianKnight]
    #26387477 - 12/17/19 04:50 PM (4 years, 1 month ago)

Quote:

StygianKnight said:
Expect to transfer to a new plate once it’s grown out a bit.




Expect to transfer because there will likely be contams present?


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:mushroom::mushroom::mushroom::mushroom::mushroom::mushroom::mushroom::mushroom::mushroom::mushroom:
:mushroom:                  :mushroom2:                :mushroom:
:mushroom:            :mushroom2::mushroom2::mushroom2:            :mushroom:
:mushroom:      :mushroom2::mushroom2::mushroom2::mushroom2::mushroom2:      :mushroom:
:mushroom::cookiemonster: BEHOLD :elmo::mushroom:
:mushroom::mushroom::mushroom::mushroom::mushroom::mushroom::mushroom::mushroom::mushroom::mushroom:


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InvisibleSpunkyMonkey88
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Re: Tissue cloning question [Re: metaphoric]
    #26387501 - 12/17/19 05:06 PM (4 years, 1 month ago)

Quote:

metaphoric said:
Quote:

StygianKnight said:
Expect to transfer to a new plate once it’s grown out a bit.




Expect to transfer because there will likely be contams present?




Ya I dunno.. I cloned three different shrooms and I don't think any of them look contaminated. So either I got lucky or I'm just  not seeing something that is there.


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InvisibleStygianKnight
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Re: Tissue cloning question [Re: metaphoric]
    #26387519 - 12/17/19 05:17 PM (4 years, 1 month ago)

Potentially, both growing and not yet growing.

Afterall agar only grows things that enjoy growing on that agar mix, it’s possible for plenty of things that grow on grain to sit on your agar or on your plate and not do anything.  So while it’s also certainly possible to put a sample from your clone agar to grain and be fine, it’s often best to get at least one plate removed from that time you touched and had a potentially dirty fruit in the environment with your plate.


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Offlinemetaphoric
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Re: Tissue cloning question [Re: StygianKnight]
    #26387567 - 12/17/19 05:39 PM (4 years, 1 month ago)

Thank you all for the info 👍

Out of curiosity do y'all what tools are you all typically using? Just a knife? Two? Tweezers?

And how are you steralizing them? Flame just before putting them in the box or PC?


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:mushroom::mushroom::mushroom::mushroom::mushroom::mushroom::mushroom::mushroom::mushroom::mushroom:
:mushroom:                  :mushroom2:                :mushroom:
:mushroom:            :mushroom2::mushroom2::mushroom2:            :mushroom:
:mushroom:      :mushroom2::mushroom2::mushroom2::mushroom2::mushroom2:      :mushroom:
:mushroom::cookiemonster: BEHOLD :elmo::mushroom:
:mushroom::mushroom::mushroom::mushroom::mushroom::mushroom::mushroom::mushroom::mushroom::mushroom:


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InvisibleSpunkyMonkey88
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Re: Tissue cloning question [Re: metaphoric]
    #26387645 - 12/17/19 06:28 PM (4 years, 1 month ago)

Pretty much just a scaple and an inoculation loop, very rarely a tweezers but I have used them.

I bought a whole dissection kit on Amazon for 8 bucks it's got everything you'll ever need minus the inoculation loop.

Flame them after any and all contact with anything


Edited by SpunkyMonkey88 (12/17/19 06:29 PM)


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