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OfflineMushroomFriend
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cloning
    #3603098 - 01/09/05 01:05 PM (11 years, 10 months ago)

Hello

I prepared an agar substitute and a jar with malt/dextrose liquid to drop a tissue of a mushroom in.
When I cut the mushroom the stem is hollow.

How to clone them?

Thanks.

MF


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Offlinemickywilliams2
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Registered: 05/27/03
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Re: cloning [Re: MushroomFriend]
    #3603117 - 01/09/05 01:12 PM (11 years, 10 months ago)

Use the cap


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OfflineMushroomFriend
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Re: cloning [Re: MushroomFriend]
    #3603126 - 01/09/05 01:14 PM (11 years, 10 months ago)

How?


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Invisiblederx
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Re: cloning [Re: MushroomFriend]
    #3603149 - 01/09/05 01:19 PM (11 years, 10 months ago)

MF, just get some of the inner mycelium from the stem, it will still work. Did you prepare peroxidated agar?


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better living through chemistry

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it's not a war on drugs, it's a war on personal freedom, ok, thats what it is.


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OfflineMushroomFriend
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Re: cloning [Re: MushroomFriend]
    #3603186 - 01/09/05 01:31 PM (11 years, 10 months ago)

no i have an agar substitute (doc34's tek) and liquid mycelium.

I just took some of the soft middle of the stem and put in the h2o2 a few seconds and it started bubbling. took it out fast and put in the liquid medium.

Cant I just slice the stem and use that?

Ah maybe I just wait till next harvest with cloning :frown:


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Invisibleblissedout
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Re: cloning [Re: MushroomFriend]
    #3603220 - 01/09/05 01:42 PM (11 years, 10 months ago)

This is an interesting question, MF. I will follow this thread, because I plan on trying this method, soon. Good luck


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:murray:


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OfflineMushroomFriend
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Re: cloning [Re: MushroomFriend]
    #3603295 - 01/09/05 02:03 PM (11 years, 10 months ago)

Thanks Blissed.

I hope for even more interesting answers! :laugh:

I miss some cloning teks (pictorials) or dont I look good enough?


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Invisiblederx
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Re: cloning [Re: MushroomFriend]
    #3603446 - 01/09/05 02:44 PM (11 years, 10 months ago)

MF, im just saying, if you're not working under a flowhood/glovebox and you're trying to clone, you should be using peroxidated agar.


--------------------
better living through chemistry

OVERGROW the government!!

it's not a war on drugs, it's a war on personal freedom, ok, thats what it is.


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OfflineMushroomFriend
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Re: cloning [Re: MushroomFriend]
    #3603618 - 01/09/05 03:28 PM (11 years, 10 months ago)

Ok, its a lowkey experiment. I dont use (peroxidated) agar.

The cardboard cloning tek HAS soem pics, but still I dont see whcih piece of the mushroom EXACTLY I must use.

Can I use a slice of the stem without spores on it?


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Edited by MushroomFriend (01/09/05 03:30 PM)


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Invisiblederx
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Re: cloning [Re: MushroomFriend]
    #3603637 - 01/09/05 03:33 PM (11 years, 10 months ago)

ya, get one of the inner pieces of the mushroom though. USE PEROXIDATED AGAR, you have to add H2O2 once the agar cools to 140 degrees. After this you wont have to worry about spore contamination.


--------------------
better living through chemistry

OVERGROW the government!!

it's not a war on drugs, it's a war on personal freedom, ok, thats what it is.


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OfflineMushroomFriend
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Re: cloning [Re: MushroomFriend]
    #3603654 - 01/09/05 03:37 PM (11 years, 10 months ago)

I dont have (PEROXIDATED) AGAR!

I can slightly "scale" the stem like a potatoe so only the inner part is left. Then there is no need to be afraid of spores anyway.

Ok I shouldnt have dipped the piece in 3% h2o2... Should be in 0,3%.. hehe :grin:


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Invisiblederx
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Re: cloning [Re: MushroomFriend]
    #3603669 - 01/09/05 03:40 PM (11 years, 10 months ago)

Ya that sounds great man, but im saying when you prepped the agar you should in future cases monitor the temperature while it's cooling and add H2O2 when it cools to 140 F. This way OTHER spores cannot contam your plates. Believe me, you will need it unless you have a flowhood/glovebox.


--------------------
better living through chemistry

OVERGROW the government!!

it's not a war on drugs, it's a war on personal freedom, ok, thats what it is.


Post Extras: Print Post  Remind Me! Notify Moderator
OfflineMushroomFriend
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Re: cloning [Re: MushroomFriend]
    #3603802 - 01/09/05 04:14 PM (11 years, 10 months ago)

I think teh cardboard tek is usefull too. And lots of things CAN be done without gloveboxes and such. Only on long run you WILL have higher contam ratio's.

But now time to tell me which part of the mushroom you use for cloning? Be as precise as possible please... :smile:

Edit: btw nice to see you (back?) again and if I am able to monitor temps in agar plates :lol: I will think bout what you said! :thumbup:


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Edited by MushroomFriend (01/09/05 04:17 PM)


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Invisibletripndicular
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Re: cloning [Re: MushroomFriend]
    #3603824 - 01/09/05 04:20 PM (11 years, 10 months ago)

The only thing that matters is that the fleash come from inside the mush somewhers and that you keep it a sterile as possible the whole time while preparing it .
Even if it is just a fibrous strand from inside the fruit body it will begin to colonize the agar . The piece only needs to be a small triangular shaped piece as big as the letter "A" on you key board is almost to big , but good size to let you know what to do .Shape is not critical , just common shape for ease of handling .
Hope this helps
GL


--------------------
Any information I give is not intended to aide you in the production of potentialy illegal substances !None of my exp comes from growing illegal varities , so take it as you will .
So with that said here is our mission statement .

Then the priest fell into a trance or swoon,& said unto the Queen of heaven ; Write unto us the ordeals; write unto us the rituals; write unto us the law !


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Invisiblederx
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Re: cloning [Re: MushroomFriend]
    #3603843 - 01/09/05 04:24 PM (11 years, 10 months ago)

i dont care if you listen or not, im just saying if you pour regular agar plates they will contam unless done in an aseptic environment. Before you pour the plates you monitor the temperature, you then add the H2O2, you know have peroxidated agar. You pour onto plates and mold and bacteria spores will not germinate on your plates.

I've never cloned a mushroom so I cannot tell you which part to take. But it seems to me, best you can do with a hollow stem is cut a small slice from the inside of the hollow stem.


--------------------
better living through chemistry

OVERGROW the government!!

it's not a war on drugs, it's a war on personal freedom, ok, thats what it is.


Post Extras: Print Post  Remind Me! Notify Moderator
Invisibletripndicular
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Re: cloning [Re: derx]
    #3603861 - 01/09/05 04:28 PM (11 years, 10 months ago)

Quote:

derx said:
Ya that sounds great man, but im saying when you prepped the agar you should in future cases monitor the temperature while it's cooling and add H2O2 when it cools to 140 F.  This way OTHER spores cannot contam your plates.  Believe me, you will need it unless you have a flowhood/glovebox.




I do not care if you pour a gallon of h2o2 into the mix , other "spores" can and will still grow . It will just "aide" iN fighting off other contams , like greens , and pin molds .
IT IS A MYTH THAT YOU MUST DO THESE TYPES OF AGAR WORK IN A GB OR HOOD !
I have done tons and with very miniaml contams . I think I have seen 6 bad dishes after pouring aprox 200+ this past year , thats good ratio IMVFO !
Maybe I am just luckiest SOB on the planet , or have too clean of a house  :wink:
But you can have great success without toys IMO . Its a matter of determination :wink:
ps never tried the h202 teks , even have the whole printing on it , just never have had need to use it yet , anyone who wants it can PM and I'll send it to you , it fills up an "A" diskett , almost .


edit...
I do reccomend that one wear a surgical mask and hair cap , and clean the livin hell outts your hands up to the elbow , and especially under then dirty booger picking finger nails  :wink: :evil:


--------------------
Any information I give is not intended to aide you in the production of potentialy illegal substances !None of my exp comes from growing illegal varities , so take it as you will .
So with that said here is our mission statement .

Then the priest fell into a trance or swoon,& said unto the Queen of heaven ; Write unto us the ordeals; write unto us the rituals; write unto us the law !


Edited by tripndicular (01/09/05 04:33 PM)


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OfflineMushroomFriend
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Re: cloning [Re: MushroomFriend]
    #3603882 - 01/09/05 04:32 PM (11 years, 10 months ago)

Yes it is helpfull Tripper! But where does the inside start? hehe
It can be done rather fast if you have sharp knife and such. Its bout the same as when I did my fake-agar to liquid in open air. As far as I know no contams yet.

But I spray the hell out of bathroom with vinagre and h2o2, use gloves often nowadays and work quick! I have a mouthcap ready to be used too.

My contams so far came from:
- bad PC-ing (wetspot)
- bad vermlayers in PF style jars. (greenmold)
- Bad casing / ventilation (cobweb)
IMHO, never know for sure but this is what I think.

Thanks a lot Tripper!

MF


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Invisiblederx
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Re: cloning [Re: tripndicular]
    #3603887 - 01/09/05 04:34 PM (11 years, 10 months ago)

i believe you are mislead and misguided and lying to the community. Don't try lying saying you do open air agar work and have only had 6 plates contam out of 200. to be blunt, you are full of it.

I'm not even going to waste my time refuting what you said, someone else can.


--------------------
better living through chemistry

OVERGROW the government!!

it's not a war on drugs, it's a war on personal freedom, ok, thats what it is.


Post Extras: Print Post  Remind Me! Notify Moderator
OfflineMushroomFriend
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Re: cloning [Re: MushroomFriend]
    #3603888 - 01/09/05 04:34 PM (11 years, 10 months ago)

lol you dont care if I listen or not, cool. I stop listening now then! :lol:

:bye:


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Invisibletripndicular
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Registered: 08/25/02
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Re: cloning [Re: MushroomFriend]
    #3603909 - 01/09/05 04:38 PM (11 years, 10 months ago)

Inside = as close to center of mush as you can get , and a decent distance from the base , inside the stem should be sterile , it has not been exposed to any air until you open and actually make the transfer .
Sterilize some tweezers in the pc , they are great for retrieving and handling small pieces .
]Does that explain what and where inside means ?

PS do work speedily , but not so fast that you stir up little whirlwinds , that could stir up cantams you missed during cleaning  :wink: Quick purposefull movements . :thumbup:


--------------------
Any information I give is not intended to aide you in the production of potentialy illegal substances !None of my exp comes from growing illegal varities , so take it as you will .
So with that said here is our mission statement .

Then the priest fell into a trance or swoon,& said unto the Queen of heaven ; Write unto us the ordeals; write unto us the rituals; write unto us the law !


Post Extras: Print Post  Remind Me! Notify Moderator
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Amazon Shop for: ½ Pint Jars, Agar, Scales

Mushrooms, Mycology and Psychedelics >> Mushroom Cultivation

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