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Offlineaccesstwo
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Registered: 04/29/06
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cloning using lc
    #5574338 - 04/30/06 02:59 PM (17 years, 9 months ago)

can you clone a mushroom by taking a piece of the inside tissue and then just dumping it in a sterilised lc jar, and let the mycillin grow.?


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OfflineCryogenicz
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Re: cloning using lc [Re: accesstwo]
    #5574345 - 04/30/06 03:01 PM (17 years, 9 months ago)

Yes.


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Offlinemikeownow
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Re: cloning using lc [Re: Cryogenicz]
    #5574423 - 04/30/06 03:30 PM (17 years, 9 months ago)

you can omg wtf I wish I knew this sooner


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Offlineebass
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Re: cloning using lc [Re: mikeownow]
    #5574434 - 04/30/06 03:34 PM (17 years, 9 months ago)

if there are spores in the tissue you can. a piece of stem will not work...how in the hell could it?


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Offlinevintage_gonzo
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Registered: 04/08/06
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Re: cloning using lc [Re: ebass]
    #5574439 - 04/30/06 03:37 PM (17 years, 9 months ago)

jesus christ this makes cloning so much easier. i thought you had to have a flow hood with agar to be anywhere near successful. What part of the mushroom would be best to cut? Just take a piece off of the bottom of a cap?


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OfflineBPJohnny
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Re: cloning using lc [Re: ebass]
    #5574465 - 04/30/06 03:45 PM (17 years, 9 months ago)

Quote:

ebass said:
if there are spores in the tissue you can. a piece of stem will not work...how in the hell could it?





cloning does not involve spores if it grows from a spore its not a
clone, so you need to be careful not to get the outside you want the
inside of the stem and try your best to do it in a way were you wont get
any spores that are very likely to be sitting on the outside of the stem


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InvisibleJaeger
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Registered: 10/01/05
Posts: 960
Re: cloning using lc [Re: vintage_gonzo]
    #5574476 - 04/30/06 03:47 PM (17 years, 9 months ago)

Take a piece from inside the stem.  Cut it in half in a glove box, and take a small piece from the center of the stipe.
Spores are not needed :smile:


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InvisibleHolydiver
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Re: cloning using lc [Re: ebass]
    #5574478 - 04/30/06 03:48 PM (17 years, 9 months ago)

Quote:

ebass said:
if there are spores in the tissue you can. a piece of stem will not work...how in the hell could it?




It's called a tissue culture, and it works 5x better than spores. Using sterile practice + tools, you want to cut a piece of flesh from inside the mushroom. Cut the cap vertically and slice a section out, or get a piece from the stem--all that really matters is that the transfer is sterile. From there you can transfer to a master grain jar, or even better, liquid culture.



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OfflineBPJohnny
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Re: cloning using lc [Re: Jaeger]
    #5574480 - 04/30/06 03:48 PM (17 years, 9 months ago)

Quote:

Jaeger said:
Take a piece from inside the stem.  Cut it in half in a glove box, and take a small piece from the center of the stipe.
Spores are not needed :smile:




:thumbup: :thumbup: :thumbup: :thumbup: :thumbup: in fact spores are not desired


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Offlineskeletor
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Re: cloning using lc [Re: BPJohnny]
    #5574485 - 04/30/06 03:51 PM (17 years, 9 months ago)

well you can't do this in open air becuase of contams of course. however you could use h202 to clean the myc. or piece of mushroom stem fleash and then stick it into an LC in a glove box or even a make shift one with a clear plastic garbage bag with an elastic around your wrist.

also i have heard of using a syringe. you wipe down the outisde of the step with alcohol then jab it with your syringe to get a little core sample. then bend it to break off that piece in the needle. then wipe the needle and injection site on your lc jar. and inject your little core sample. this i think would be the best for open air.

ive yet to try it but i will next week when i have something to try it on.


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OfflineP.Menace
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Re: cloning using lc [Re: skeletor]
    #7747952 - 12/12/07 02:53 AM (16 years, 2 months ago)

Quote:

skeletor said:
also i have heard of using a syringe. you wipe down the outisde of the step with alcohol then jab it with your syringe to get a little core sample. then bend it to break off that piece in the needle. then wipe the needle and injection site on your lc jar. and inject your little core sample. this i think would be the best for open air.




This is the best way to do it. as long as your needles are sterile... one could use a clean sterile needle, suck up a small ammount 1/4cc or so H202, make the "Core Sample", dunk the entire needle (not the whole syringe) in H202 after the sample has been taken, then inject the whole solution and tissue sample into an LC.
The H202 will not harm the myc. or the LC. I have seen myc. grow out in nothing but H202, because it degrades over time turning into water


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Offlineveda_sticks
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Re: cloning using lc [Re: P.Menace]
    #7748135 - 12/12/07 06:34 AM (16 years, 2 months ago)

Quote:

P.Menace said:
Quote:

skeletor said:
also i have heard of using a syringe. you wipe down the outisde of the step with alcohol then jab it with your syringe to get a little core sample. then bend it to break off that piece in the needle. then wipe the needle and injection site on your lc jar. and inject your little core sample. this i think would be the best for open air.




This is the best way to do it. as long as your needles are sterile... one could use a clean sterile needle, suck up a small ammount 1/4cc or so H202, make the "Core Sample", dunk the entire needle (not the whole syringe) in H202 after the sample has been taken, then inject the whole solution and tissue sample into an LC.
The H202 will not harm the myc. or the LC. I have seen myc. grow out in nothing but H202, because it degrades over time turning into water




thats not quite true about h202, while it degrades over time, and that the myc may have been able to grow in it. Its not quite true that h202 does not harm myc, infact h202 is more harmfull to myc than contams (h202 is only usefull against cobweb, its not effective against anything else)

If you are having problems with contaminated LC's from cloning, you can try soaking the tissue in a bleach solution for a few minutes. This will do less damage than h202 and i believe its much better at cleaning the sample up.

Ofcoarse cloning to agar would be much better.


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Invisibleuncle_rico
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Re: cloning using lc [Re: veda_sticks]
    #7748164 - 12/12/07 06:54 AM (16 years, 2 months ago)

Is there a tek or post that discusses h2o2 vs bleach water for 'core biopsy to LC' cloning ?

I was planning on using sterile water but any additional layer of security is welcomed.


I think this 4" long, 14 gauge needle will core a nice biopsy !

These are $ .50 at a local feed store.


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OfflineNibin
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Re: cloning using lc [Re: P.Menace]
    #7748252 - 12/12/07 07:45 AM (16 years, 2 months ago)

http://www.shroomery.org/forums/showflat.php?Cat=0&Board=2&Number=5874305&fpart=&PHPSESSID=

A very informative tek on using LC's to grow a tissue sample obtained with a needle.


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Invisibleuncle_rico
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Re: cloning using lc [Re: Nibin]
    #7748284 - 12/12/07 08:00 AM (16 years, 2 months ago)

Excellent. Thanks


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