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OfflineCloneufc
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Cloneufc's easy Cloning tek * 9
    #12032286 - 02/15/10 06:13 AM (13 years, 11 months ago)

This is a simple cloning tek that has worked well for me. It can be used with agar,without agar or without antibiotic agar. This cloning tek can also be used to make an LC. This tek has been edited to help people from making some mistakes. Things you'll need: A scapel or razor knife,an unopened ziploc baggie. You should have everything else.

I have had trouble with most of the cloning teks on here. I've tried dropping core tissue into LC jars and I would either get bacteria or no growth at all. I tried using the tek where you rub down the mushroom stem with alcohol and poke it through with a syringe. I would not get any tissue in the needle, the needles we use in this hobby were made to not rip chunks out of living tissue. I then used a modified blunt tip needle to get tissue and inject it into an LC. Same thing bacteria or no growth at all. Then I tried agar with core tissue, it was hit or miss trying to dodge bacteria. Bacteria is a lot faster and would turn the core tissue into goop.


Heres how you do it.
1. Turn on your flowhood or prep your glovebox.
2. Take the mushroom you want to clone and tear it in half    infront of your flowhood or inside your glovebox.(Use gloves, rubbing alcohol on them.)
3. Heat your scalpel red hot and cut out a small piece of tissue from inside the stem base. I like to use the stem instead of the cap to lesson the chance of getting spores on my tissue.
4. While working in front or inside your glovebox take an unopened zip lock baggie and place the tissue inside of it and zip it up. Flatten it out to get most of the air out. Label the ziploc bag with a sharpie. The purpose of this is to keep the tissue in a nonnutrient enviroment. That way it doesnt have to compete with bacteria or other molds. It also keeps the tissue hydrated and gives the tissue-mycelium a big head start. After about 3-6 days the tissue will become a puff ball of mycelium. What surprised me, is that I have not had any tissue rot issues so far.

(Do NOT dip the tissue in hydrogen peroxide, it will kill it or weaken it. When placing your tissue in the ziploc bag make sure it is on the dry side. You want the tissue slightly moist but not wet. It will recover faster this way. If the tissue is wet it will start rotting.)



Go here if your using agar:
5. Now transfer the tissue from the ziploc bag to your agar. Now its ready to do what you want with it.(Use sterile tweezers or sterile scalpel to transfer, while trying not to damage the mycelium.)

Go here if you dont use or have agar:
5.Now transfer the tissue from the ziploc bag to a grain jar. Once colonized you can make grain transfers or a grain LC..(Use sterile tweezers or sterile scalpel to transfer, while trying not to damage the mycelium.)


Sorry for the terrible pics but my good camera got stolen.

Here you can see the mycelium growing after a couple days.


Here's cloned tissue to agar after a couple days.


Edited by Cloneufc (02/22/10 05:10 AM)


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Invisiblebare.whiterabbit
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Registered: 11/29/09
Posts: 912
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Re: Cloneufc's easy Cloning tek [Re: Cloneufc]
    #12032294 - 02/15/10 06:18 AM (13 years, 11 months ago)

You've inspired me to do a hell of a lot of transfers now


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InvisibleFleadh
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Registered: 05/11/09
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Re: Cloneufc's easy Cloning tek [Re: bare.whiterabbit]
    #12032312 - 02/15/10 06:27 AM (13 years, 11 months ago)

nice, book marked


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OfflineCloneufc
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Re: Cloneufc's easy Cloning tek [Re: Fleadh]
    #12032360 - 02/15/10 06:54 AM (13 years, 11 months ago)

Im just trying to help out everyone. This is really helpful for people who dont know how to use agar.


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OfflineSanguine
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Re: Cloneufc's easy Cloning tek [Re: Cloneufc]
    #12032480 - 02/15/10 07:57 AM (13 years, 11 months ago)

:thumbup::thumbup::thumbup:

This is great. It totally confirms some things I was guessing but hadn't tried yet. Now that I'm doing agar I will definitely do this when I clone my next specimen.


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"The greatest delight which the fields and woods minister is the suggestion of an occult relation between man and the vegetable. I am not alone and unacknowledged. They nod to me, and I to them." --Ralph Waldo Emerson

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InvisibleLeopardMan
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Registered: 09/21/09
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Re: Cloneufc's easy Cloning tek [Re: Sanguine]
    #12032500 - 02/15/10 08:08 AM (13 years, 11 months ago)

Will this work even if I use heatshocked mushrooms? :lol: I am joking, congrats. :congrats:


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You have to die a few times before you can really live.

-Charles Bukowski-


Edited by LeopardMan (02/15/10 08:08 AM)


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Invisiblemyceleus_rex
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Re: Cloneufc's easy Cloning tek [Re: LeopardMan]
    #12032637 - 02/15/10 09:00 AM (13 years, 11 months ago)

"Will this work even if I use heatshocked mushrooms?" :rofl2:

This is way cool :cool: Thanks!


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Cervantes' sclerotia thread    Citric's self-healing lid tek  Agar's Grain LC  Breaking Up Spawn Jars

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InvisibleJonEveryman88
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Re: Cloneufc's easy Cloning tek [Re: myceleus_rex]
    #12032798 - 02/15/10 09:46 AM (13 years, 11 months ago)

Hm. Cool I'm gonna try this right now.

Nice tek:thumbup:


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Invisibleprismism
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Registered: 05/11/09
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Re: Cloneufc's easy Cloning tek [Re: Cloneufc]
    #12032869 - 02/15/10 10:01 AM (13 years, 11 months ago)

i've never thought to use ziploc bags. i always get bacteria when trying to make clone lcs.. so i shall give this a whirl.


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OfflineKillaFoRilla
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Re: Cloneufc's easy Cloning tek [Re: prismism]
    #12033018 - 02/15/10 10:38 AM (13 years, 11 months ago)

The idea of waiting for new healthy myc is pretty spiffy. Gonna give this a try next time.


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Invisibletruskool
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Re: Cloneufc's easy Cloning tek [Re: KillaFoRilla]
    #12033053 - 02/15/10 10:44 AM (13 years, 11 months ago)

:thumbup:
another great tek by clone your a great addition to the community


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Invisiblebadman
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Re: Cloneufc's easy Cloning tek [Re: truskool]
    #12033095 - 02/15/10 10:55 AM (13 years, 11 months ago)

Two thumbs up
:awethumb::awethumb:

:superiority:


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OfflineMissShroom
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Registered: 10/31/09
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Re: Cloneufc's easy Cloning tek [Re: badman]
    #12033507 - 02/15/10 12:09 PM (13 years, 11 months ago)

Thanks! I've also had the no-grow -problem sometimes when cloning, gonna try this one next time :thumbup:.


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OfflineCloneufc
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Re: Cloneufc's easy Cloning tek [Re: MissShroom]
    #12035000 - 02/15/10 04:06 PM (13 years, 11 months ago)

Ive read so many times that indoor mushrooms dont carry bacteria so antibiotic agar is not needed. Well for me and countless others indoor mushrooms do carry bacteria, maybe not as much. Im pretty sure this tek will work with wild clones. I know that indoor mushrooms do carry bacteria because on agar the bacteria only grows where that mushroom touched the agar. With cloning, Ive never struggled with molds only bacteria. With this tek the mycelium comes out of the ziploc bag with little to no bacteria.


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Invisibleblood4blood
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Registered: 04/25/07
Posts: 6,029
Loc: The Valley
Re: Cloneufc's easy Cloning tek [Re: Cloneufc]
    #12035223 - 02/15/10 04:42 PM (13 years, 11 months ago)

good write up clone! you need to work on those pics though my friend.:tongue:


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Invisibleanonjon
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Posts: 6,322
Re: Cloneufc's easy Cloning tek [Re: blood4blood]
    #12035328 - 02/15/10 05:05 PM (13 years, 11 months ago)

That's interesting how you leave it in the ziplock to turn back into myc first.

Something you might want to try sometime is dipping tissue in straight h202 for 10-30 seconds before doing wutever with it. Works like a charm.


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The above post is fictional, hypothetical, or downright nonsensical.


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Invisiblemyceleus_rex
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Re: Cloneufc's easy Cloning tek [Re: anonjon]
    #12035464 - 02/15/10 05:29 PM (13 years, 11 months ago)

By "straight" you must mean 3% off the shelf stuff. Real "straight" H2O2 would turn myc to mush in no time.


--------------------
Cervantes' sclerotia thread    Citric's self-healing lid tek  Agar's Grain LC  Breaking Up Spawn Jars

World's Ugliest Cat?

If ignorance is bliss, why aren't there more happy people?


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Invisibletruskool
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Re: Cloneufc's easy Cloning tek [Re: myceleus_rex]
    #12035473 - 02/15/10 05:31 PM (13 years, 11 months ago)

Yes 3% pure would fuck up your tissue.  It does work pretty good tho IME


--------------------
Ask  AMU    for    the  best    mycology    advice  out  there

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Invisiblemyceleus_rex
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Re: Cloneufc's easy Cloning tek [Re: truskool]
    #12035703 - 02/15/10 06:13 PM (13 years, 11 months ago)

Quote:

truskool said:
Yes  3% pure would fuck up your tissue.  It does work pretty good tho IME



3% pure sounds OK, it's "pure", as in 100% that I was talking about.


--------------------
Cervantes' sclerotia thread    Citric's self-healing lid tek  Agar's Grain LC  Breaking Up Spawn Jars

World's Ugliest Cat?

If ignorance is bliss, why aren't there more happy people?


Extras: Filter Print Post Top
Invisibletruskool
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Registered: 06/28/09
Posts: 11,194
Loc: Over the rainbow
Re: Cloneufc's easy Cloning tek [Re: myceleus_rex]
    #12035713 - 02/15/10 06:15 PM (13 years, 11 months ago)

thats wat I was saying sorry if im the king of run on sentences or dont make myself clear enough:crazy2:


--------------------
Ask  AMU    for    the  best    mycology    advice  out  there

  Roll it :burnone:While I troll it.  :maximumtrolling:

I don't mean to boast, but dam if I don't brag.


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