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OfflineVibration
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Cloning a mushroom
    #14132461 - 03/16/11 05:44 PM (12 years, 10 months ago)

http://www.shroomery.org/forums/showflat.php/Number/5874305#5874305

Quote:

Quickly wipe your needle with iso (flame if desired, I don't bother - we did just sterilize it after all!) and stab it straight through the stem to the other side.



Quote:

Give the needle another wipe, insert into hole, and push the plunger.  The water in the syringe will force the cross section of the stem out into your LC.




I'm a bit confused. When stabbing the mushroom with a syringe, will the tissue simply get into the needle.. or wtf? .. so when I inject it into my LC jar, it would drop in?

Would it work on a jar with a self-healing injection port?

And one more thing. I've harvested my mushroom for about 21 hours ago, I've kept it in a cool room with a temperature of 10C in a isolated bag, is it still good to use for cloning process??


Many thanks for solving my confusion!


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Offlineesquaredx
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Re: Cloning a mushroom [Re: Vibration]
    #14132546 - 03/16/11 06:00 PM (12 years, 10 months ago)

The chunk of mushroom will get lodged in your needle.
And yes, it will work with a self healing port..
Just push that wad into your LC and be good.

Idk if I would use mushrooms that have been sitting out. Im sure you could tho if you were really sterile.
Slice the mushroom and take some tissue from the inside :shrug:


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Invisibleabltsandwich
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Re: Cloning a mushroom [Re: Vibration]
    #14132560 - 03/16/11 06:02 PM (12 years, 10 months ago)

The idea is that the needle takes a core sample, so to speak, of the shroom and then when you press the plunger the core sample goes out into the LC.  Yes, a self-healing RTV port will be fine.  Your shroom should be fine for cloning.

That said, I would recommend not going with that tek.  It's given nothing but headaches, since it's so hard to get the needle to get a core sample.  Plus dropping a core sample straight into LC isn't the best route.  Drop your sample into some grains, let that colonize, and make a grain LC, from which you can make working LC in whatever amount you like.


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Offlineesquaredx
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Re: Cloning a mushroom [Re: abltsandwich]
    #14132572 - 03/16/11 06:04 PM (12 years, 10 months ago)

I would just keep that Grain LC and be happy.
No cores needed. Just a nice jar of grains and some clean water :thumbup:


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Invisiblebiologys
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Re: Cloning a mushroom [Re: esquaredx]
    #14132595 - 03/16/11 06:08 PM (12 years, 10 months ago)

i've never had any luck getting any tissue in a needle as well, i just put the fruit in a glovebox, have sterile tweezers, rip the fruit in half and take a small section with tweezers then drop on agar, or in your case LC


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OfflineHybridprX
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Re: Cloning a mushroom [Re: esquaredx]
    #14132621 - 03/16/11 06:12 PM (12 years, 10 months ago)

Whoever posted that tek must be a magician for it to have worked, simply stabbing a mushroom stem and injecting a liquid culture or agar is almost certain to doom.

In order to clone you are definitely going to require a glovebox and the tissue should be taken from the base of the stem from the largest cluster...cleaned on agar and then propagated to a liquid culture.

Another way is to cutt a small square from the base of the mushroom, dip it h202 solution and then propagate it to a LC or a grain jar...

Long story short, build a glovebox or buy a flowhood.


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Invisiblemonotubgirl
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Re: Cloning a mushroom [Re: Vibration]
    #14132881 - 03/16/11 06:58 PM (12 years, 10 months ago)

well hello there and here's what i think

cloning out a shroom
take the tissue from the shroom
put it on agar



Edited by monotubgirl (03/16/11 07:04 PM)


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OfflineDamion5050
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Re: Cloning a mushroom [Re: monotubgirl]
    #14132908 - 03/16/11 07:02 PM (12 years, 10 months ago)

Its simple..

1 Prepare some petri dishes.

2 Take your mushroom and swab it with some cotton balls soaked in 70% alcohol

3 Take a sterile scalpel and cut the shroom down the middle to expose the inner tissue.

4 Re sterilize your scalpel and cut a chunk of the inner tissue out

5 Dunk your tissue in a shot glass filled with peroxide..  (about 20-30 seconds)

6 Place tissue sample on agar dish and close it up..


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OfflineVibration
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Re: Cloning a mushroom [Re: monotubgirl]
    #14132961 - 03/16/11 07:14 PM (12 years, 10 months ago)

Thank you very much for the advice, but I'll better stick to my LC as I will need to study agar first and I don't have enough time for it at the moment. Also my LC is waiting for me already. Mushroom too!

I just have to go with a LC this time. I promise I will learn to do AGAR soon.

I'm going to try to get the tissue with a help of my glovebox somehow. With a needle or tweezers. Damn it, I wish I had a scalpel available :laugh:


(I'm will post later what I did and then I'll post the results too so you can see what turned out)


--------------------
The trick is to combine your waking rational abilities with the infinite possibilities of your dreams. Because, if you can do that, you can do anything.


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OfflineDamion5050
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Re: Cloning a mushroom [Re: Vibration]
    #14133032 - 03/16/11 07:25 PM (12 years, 10 months ago)

You can also drop the tissue sample into an LC, but the thing with agar is you can make sure there are no contaminates..  Where if you drop it in an LC then wait for the LC to colonize then inject your LC into some jars then your jars go south you have just waisted a few weeks of work..


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InvisibleDoc_T
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Re: Cloning a mushroom [Re: Damion5050]
    #14133058 - 03/16/11 07:28 PM (12 years, 10 months ago)

I never did get tissue->LC to work.


--------------------
You make it all possible. Doesn't it feel good?


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OfflineDamion5050
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Re: Cloning a mushroom [Re: Doc_T]
    #14133063 - 03/16/11 07:28 PM (12 years, 10 months ago)

Quote:

Doc_T said:
I never did get tissue->LC to work.





I have gotten it to work but prefer to do agar just to make sure there are no contams..


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OfflineVibration
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Re: Cloning a mushroom [Re: Damion5050]
    #14133145 - 03/16/11 07:40 PM (12 years, 10 months ago)

I have 5 jars of honey/water solution.

If "tissue>LC" really is so bad, then I will just use 1 jar for the attempt to clone my first and big fast grown mushroom, which have been waiting in a cold room for 24 hours already. We can see how did it turn out in a few weeks.

Other 4 jars I will use with a regular "spore>LC" method then.


I really think it's the best I could do right now. Although, I will keep that in mind what you've told me.


--------------------
The trick is to combine your waking rational abilities with the infinite possibilities of your dreams. Because, if you can do that, you can do anything.


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OfflineDamion5050
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Re: Cloning a mushroom [Re: Vibration]
    #14133159 - 03/16/11 07:43 PM (12 years, 10 months ago)

Quote:

Vibration said:
I have 5 jars of honey/water solution.

If "tissue>LC" really is so bad, then I will just use 1 jar for the attempt to clone my first and big fast grown mushroom, which have been waiting in a cold room for 24 hours already. We can see how did it turn out in a few weeks.

Other 4 jars I will use with a regular "spore>LC" method then.


I really think it's the best I could do right now. Although, I will keep that in mind what you've told me.





By no means am I saying LC is bad..  But when you are taking a clone, you are taking tissue from something that isn't sterile and you are putting it into a LC which is sugar based and contams will love it and eat the hell out of it and next thing u know your LC is bad and your no wiser to it until you knock up your jars..  I love using LC but I always tend to go tissue to agar then agar to LC..  And I even do spores to agar then agar to LC..


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OfflineVibration
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Re: Cloning a mushroom [Re: Damion5050]
    #14133230 - 03/16/11 07:58 PM (12 years, 10 months ago)

Really seems like I will have to start doing research about agar. Damn it :grin:


--------------------
The trick is to combine your waking rational abilities with the infinite possibilities of your dreams. Because, if you can do that, you can do anything.


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OfflineDamion5050
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Re: Cloning a mushroom [Re: Vibration]
    #14133242 - 03/16/11 07:59 PM (12 years, 10 months ago)

Agar is easy, you can make it from scratch or buy some pre mixed..  I bought premixed, all you do is add water and PC then pour your dishes in front of a flowhood or in glove box..


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Invisiblebiologys
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Re: Cloning a mushroom [Re: Damion5050]
    #14133264 - 03/16/11 08:02 PM (12 years, 10 months ago)

Quote:

Damion5050 said:
Agar is easy, you can make it from scratch or buy some pre mixed..  I bought premixed, all you do is add water and PC then pour your dishes in front of a flowhood or in glove box..





i've been telling him this for few weeks now


honestly man, agaris super simple once you do it for your first time you're going to smack yourself for not learning it sooner


i myself for the first attempt started out with the grocery store agar tek, and it worked perfectly fine..now im using LME and doing good as well.


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Offlinehamloaf
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Re: Cloning a mushroom [Re: Damion5050]
    #14133269 - 03/16/11 08:02 PM (12 years, 10 months ago)

Quote:

Damion5050 said:
Quote:

Vibration said:
I have 5 jars of honey/water solution.

If "tissue>LC" really is so bad, then I will just use 1 jar for the attempt to clone my first and big fast grown mushroom, which have been waiting in a cold room for 24 hours already. We can see how did it turn out in a few weeks.

Other 4 jars I will use with a regular "spore>LC" method then.


I really think it's the best I could do right now. Although, I will keep that in mind what you've told me.





By no means am I saying LC is bad..  But when you are taking a clone, you are taking tissue from something that isn't sterile and you are putting it into a LC which is sugar based and contams will love it and eat the hell out of it and next thing u know your LC is bad and your no wiser to it until you knock up your jars..  I love using LC but I always tend to go tissue to agar then agar to LC..  And I even do spores to agar then agar to LC..



Why not just cut out the LC step and take your clone culture on agar to sterilized grains expanding your mycelium with grain to grain transfers?


--------------------

   
How I Get Stuff done. - My Reference Guide. - My Grows.


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Invisiblebiologys
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Re: Cloning a mushroom [Re: hamloaf]
    #14133282 - 03/16/11 08:05 PM (12 years, 10 months ago)

dont add extra steps ham, we're finally getting him talked into going to agar....baby steps, you start adding g2g you're gonna scare him again :lmafo:


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OfflineDamion5050
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Re: Cloning a mushroom [Re: hamloaf]
    #14133312 - 03/16/11 08:11 PM (12 years, 10 months ago)

Quote:

hamloaf said:
Quote:

Damion5050 said:
Quote:

Vibration said:
I have 5 jars of honey/water solution.

If "tissue>LC" really is so bad, then I will just use 1 jar for the attempt to clone my first and big fast grown mushroom, which have been waiting in a cold room for 24 hours already. We can see how did it turn out in a few weeks.

Other 4 jars I will use with a regular "spore>LC" method then.


I really think it's the best I could do right now. Although, I will keep that in mind what you've told me.





By no means am I saying LC is bad..  But when you are taking a clone, you are taking tissue from something that isn't sterile and you are putting it into a LC which is sugar based and contams will love it and eat the hell out of it and next thing u know your LC is bad and your no wiser to it until you knock up your jars..  I love using LC but I always tend to go tissue to agar then agar to LC..  And I even do spores to agar then agar to LC..



Why not just cut out the LC step and take your clone culture on agar to sterilized grains expanding your mycelium with grain to grain transfers?





I do that as well..  Just depends on what im trying to do..  I use LC to do bags and grain to grain to do jars for mono's


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