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Offlinebillycorgan55
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Registered: 11/14/08
Posts: 537
Last seen: 11 years, 9 months
Cloning with a pin.
    #16123578 - 04/22/12 08:50 AM (11 years, 9 months ago)

Anyone had success with dropping a pin into a sterilized grain jar?

I have had failure with LCs and tissue transfers. This seems like it would work if I rubbed the outside of the pin with alchohol.

Any input?


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OM MADNE PADME OBIWAN LUKE HUM....


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Offlineuninc4life2010
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Re: Cloning with a pin. [Re: billycorgan55]
    #16123592 - 04/22/12 08:56 AM (11 years, 9 months ago)

This works great on an agar plate!  RR had a chapter about this in his DVD.  Agar seems to be the way to go.  Accordin to his lesson, the mycelium grows so fast at that stage that it will outgrow any contams and sterilizing isn't necessary.


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OfflineRogerRabbitM
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Re: Cloning with a pin. [Re: billycorgan55]
    #16123598 - 04/22/12 08:59 AM (11 years, 9 months ago)

Quote:

billycorgan55 said:
This seems like it would work if I rubbed the outside of the pin with alchohol.

Any input?




Rubbing the pin in or with alcohol will ensure failure.  Place the pin on agar, let it grow for a few days, and then take tiny samples of mycelium from the leading edge of the growth to new plates to grow out and use.
RR

  -----> 


--------------------
Download Let's Grow Mushrooms



semper in excretia sumus solim profundum variat

"I've never had a failed experiment.  I've only discovered 10,000 methods which do not work."
Thomas Edison


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Offlinevvitchdoctor
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Re: Cloning with a pin. [Re: RogerRabbit]
    #16123651 - 04/22/12 09:23 AM (11 years, 9 months ago)

dipping it in H202 works well also.  I sterilize soup cans of substrate with tinfoil caps, then cut the stem butts off of my best mushrooms and set them on top - works great.  I use the stem butt/agar tek for G2G or WBS, but to keep my mycelium running I just clone out.


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OfflineRogerRabbitM
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Re: Cloning with a pin. [Re: vvitchdoctor]
    #16123663 - 04/22/12 09:27 AM (11 years, 9 months ago)

The pictures above were done with no peroxide.  Peroxide is toxic to mushroom mycelium, and when you're trying to clone, don't do anything to shock or damage the mycelium.  Simply place the pin on agar, let it grow, and if contaminants show up, transfer away from them.  That's the purpose of agar, not to prevent contaminants from growing in the first place-we want them to.
RR


--------------------
Download Let's Grow Mushrooms



semper in excretia sumus solim profundum variat

"I've never had a failed experiment.  I've only discovered 10,000 methods which do not work."
Thomas Edison


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Offlinevvitchdoctor
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Re: Cloning with a pin. [Re: RogerRabbit]
    #16123686 - 04/22/12 09:37 AM (11 years, 9 months ago)

wow!  thanks for that - I haven't had a problem with it, but I didn't know - I don't want to spread bad info.  Appreciate the education!  Will change my tek accordingly.


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Offlinebillycorgan55
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Re: Cloning with a pin. [Re: RogerRabbit]
    #16123994 - 04/22/12 11:12 AM (11 years, 9 months ago)

Quote:

RogerRabbit said:
Quote:

billycorgan55 said:
This seems like it would work if I rubbed the outside of the pin with alchohol.

Any input?




Rubbing the pin in or with alcohol will ensure failure.  Place the pin on agar, let it grow for a few days, and then take tiny samples of mycelium from the leading edge of the growth to new plates to grow out and use.
RR

  -----> 




Will each transfer taken from the pin on agar be a guarenteed fruiting strain?

My guess is no because of the mixed genetics that can be present in one mushroom. (My pins are from a MS)


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OM MADNE PADME OBIWAN LUKE HUM....


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OfflineChez man
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Re: Cloning with a pin. [Re: billycorgan55]
    #16124015 - 04/22/12 11:19 AM (11 years, 9 months ago)

Is there criteria for chosing the best pin candidates?

I'd suspect the best are the meatest, healthiest pins that first appear.


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http://www.hedweb.com/hedab.htm


Edited by Chez man (04/22/12 11:19 AM)


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InvisibleStygianKnight
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Re: Cloning with a pin. [Re: Chez man]
    #16124053 - 04/22/12 11:29 AM (11 years, 9 months ago)

My first clone I thought "I'll be extra sterile by whipping it with alcohol."  Not only has that been my only failure, but ironically it was my first contamination too.  Don't do that.


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OfflineRogerRabbitM
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Re: Cloning with a pin. [Re: billycorgan55]
    #16125325 - 04/22/12 05:51 PM (11 years, 9 months ago)

Quote:

billycorgan55 said:
Quote:

RogerRabbit said:
Quote:

billycorgan55 said:
This seems like it would work if I rubbed the outside of the pin with alchohol.

Any input?




Rubbing the pin in or with alcohol will ensure failure.  Place the pin on agar, let it grow for a few days, and then take tiny samples of mycelium from the leading edge of the growth to new plates to grow out and use.
RR

  -----> 




Will each transfer taken from the pin on agar be a guarenteed fruiting strain?

My guess is no because of the mixed genetics that can be present in one mushroom. (My pins are from a MS)




Correct, unless there's no sectoring.  Note in the petri dish section at 12 O'clock there is at least 5 strains(sectors).  Each of these strains would need to be grown out to find the best one.  The bottom left section appears to be an isolate, and the bottom right culture has two sectors that I can see right off.(7 to 9 O'clock appears to be different mycelium)
RR


--------------------
Download Let's Grow Mushrooms



semper in excretia sumus solim profundum variat

"I've never had a failed experiment.  I've only discovered 10,000 methods which do not work."
Thomas Edison


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Offlinebillycorgan55
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Registered: 11/14/08
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Re: Cloning with a pin. [Re: RogerRabbit]
    #16125670 - 04/22/12 07:12 PM (11 years, 9 months ago)

Thank you, I will begin working with agar soon.

Wish me luck!


--------------------
OM MADNE PADME OBIWAN LUKE HUM....


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Offlineelkart
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Re: Cloning with a pin. [Re: billycorgan55]
    #16125899 - 04/22/12 08:07 PM (11 years, 9 months ago)

I don't quite understand. So you'd have to do a small test grow to see how each pin performs? I had thought that you'd take the strongest fruit to make clones?


--------------------
Shroom Honey Preservation Tek


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Offlinevvitchdoctor
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Registered: 05/16/11
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Re: Cloning with a pin. [Re: elkart]
    #16126538 - 04/22/12 10:33 PM (11 years, 9 months ago)

I have a question as well - if peroxide is damaging to mycelium, why use it to fend off contaminants?  I am not trying to sharp shoot you, just trying to clear up some confusion by asking the expert... little help?


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Offlineelkart
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Re: Cloning with a pin. [Re: vvitchdoctor]
    #16127780 - 04/23/12 09:34 AM (11 years, 9 months ago)

Quote:

vvitchdoctor said:
I have a question as well - if peroxide is damaging to mycelium, why use it to fend off contaminants?  I am not trying to sharp shoot you, just trying to clear up some confusion by asking the expert... little help?




From what I understand, it's sort of a last ditch effort to save things. I recall RR saying saying something about it killing contam but nearly fatally damaging mycelium in the process, but that it would recover


--------------------
Shroom Honey Preservation Tek


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OfflineRogerRabbitM
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Re: Cloning with a pin. [Re: vvitchdoctor]
    #16127950 - 04/23/12 10:24 AM (11 years, 9 months ago)

Quote:

vvitchdoctor said:
I have a question as well - if peroxide is damaging to mycelium, why use it to fend off contaminants?




You don't. 

The only time I would ever use peroxide is if there's a cobweb outbreak on a cased bulk substrate.  The peroxide will melt the cobweb mold on contact.  It also damages the mushroom mycelium it touches, but the cobweb would destroy it outright if left unchecked.

Peroxide is useless on any other contaminant during the growing process.
RR


--------------------
Download Let's Grow Mushrooms



semper in excretia sumus solim profundum variat

"I've never had a failed experiment.  I've only discovered 10,000 methods which do not work."
Thomas Edison


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Offlinebillycorgan55
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Registered: 11/14/08
Posts: 537
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Re: Cloning with a pin. [Re: elkart]
    #16127979 - 04/23/12 10:33 AM (11 years, 9 months ago)

Quote:

elkart said:
I don't quite understand. So you'd have to do a small test grow to see how each pin performs? I had thought that you'd take the strongest fruit to make clones?




Cloning and Isolating are two different things.


--------------------
OM MADNE PADME OBIWAN LUKE HUM....


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OfflineDjchrist
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Re: Cloning with a pin. [Re: billycorgan55]
    #16128191 - 04/23/12 11:41 AM (11 years, 9 months ago)

Very useful information. Thank you


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OfflineSynapse Trap
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Re: Cloning with a pin. [Re: Djchrist]
    #16128304 - 04/23/12 12:16 PM (11 years, 9 months ago)

Nothing to add, except when I read the title of your thread, it makes me want to take a couple clonapin.. too bad I don't have any. Thanks a lot!
:loldongs:


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Synapse Trap

:flyhigh:


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Invisiblechuptoo
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Registered: 11/07/11
Posts: 523
Re: Cloning with a pin. [Re: Synapse Trap]
    #16128515 - 04/23/12 01:09 PM (11 years, 9 months ago)

its klonopin.  and dont eat pharmies unless you have to.  :smile:


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OfflineSynapse Trap
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Re: Cloning with a pin. [Re: chuptoo]
    #16128525 - 04/23/12 01:11 PM (11 years, 9 months ago)

Quote:

chuptoo said:
its klonopin.  and dont eat pharmies unless you have to.  :smile:




yeah that.. and I'll take all the ones you don't want.

:thedude:


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Synapse Trap

:flyhigh:


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