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OfflineActionjunkie
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Cloning Tek, best way to multiply?
    #26449659 - 01/24/20 09:42 AM (4 years, 5 days ago)

I've dabbled a little with cloning. The contamination rate was high. I was told that normal. So I was going to:

1. Make several Petri dishes from one sample
2. Transfer a clean mycillium sample, to fresh agar as soon as I see it
3. Take clean culture and complete agar to agar transfers to multiply
4. Inoculate grain with clean culture

I feel like I'm missing something? Any help, advice, experience would be awesome.

Thanks in advance

J+


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Consuming mushrooms may not be addictive but growing them is!


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OfflineHuskies
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Re: Cloning Tek, best way to multiply? [Re: Actionjunkie]
    #26449671 - 01/24/20 09:49 AM (4 years, 5 days ago)

That is the jist of it, just make sure to do 3-4 transfer to ensure a clean sample.

Don't get the first exact growth, but let it sector out, and pick the strongest/fastest growing sector. If it's from a clone, it will provide fruits I'm pretty sure.

Mold is the biggest killer, make sure you have at least a still air box, if you have the space, maybe just try building a flowhood, should be around 200 buckarinos if DIY


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I call them Huskies cause you tell them to go "Mush! Mush""


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InvisibleGoombas69
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Registered: 12/15/19
Posts: 15
Re: Cloning Tek, best way to multiply? [Re: Actionjunkie]
    #26449691 - 01/24/20 10:04 AM (4 years, 5 days ago)

I try to be overly cautious so I will make a backup LC at the start and end of the process so I have unlimited mycelium to work with. Then I agar the LC before I use it to get an indication if it’s clean or not. I have syringe filters in my LC just in case it is dirty and I need to try some hydrogen peroxide cleansing. I understand you can clean on agar too. Hopefully I never have to deal with that. I just don’t want to ever have to rebuy duplicate spores because of contam.


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OfflineA.k.aM
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Re: Cloning Tek, best way to multiply? [Re: Goombas69]
    #26449694 - 01/24/20 10:07 AM (4 years, 5 days ago)

Wouldn’t making spore prints be way easier??

With the cloning once you get the method down there shouldn’t be a lot of contamination.

I had a LOT of bacteria my first few but since then they’ve been clean.

Tearing it open without touching the inside and then cutting a piece out without touching the outside will usually do the trick.


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LAGM2020


Edited by A.k.a (01/24/20 10:08 AM)


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OfflineActionjunkie
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Re: Cloning Tek, best way to multiply? [Re: A.k.a] * 1
    #26449811 - 01/24/20 11:38 AM (4 years, 5 days ago)

Wouldn't a spore print just be a genetic grab bag, like a Ms syringe? Instead of being a clone?

My success rate working with spores is really good this last run. I was already planning on making a spore print and grabbing clone tissue from the stem.

And btw, you guy fucking rock!!! I love this site!!! Thank you everyone that contributes!!!

J+


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Offlinefeldman114
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Re: Cloning Tek, best way to multiply? [Re: Goombas69]
    #26449819 - 01/24/20 11:44 AM (4 years, 5 days ago)

Quote:

Goombas69 said:
I try to be overly cautious so I will make a backup LC at the start and end of the process so I have unlimited mycelium to work with. Then I agar the LC before I use it to get an indication if it’s clean or not. I have syringe filters in my LC just in case it is dirty and I need to try some hydrogen peroxide cleansing. I understand you can clean on agar too. Hopefully I never have to deal with that. I just don’t want to ever have to rebuy duplicate spores because of contam.



Full stop.
Peroxide can’t be used to clean anything. Also, how would you “clean” an LC? With agar, the term refers to making transfers of transfers from transfers, until you’re sure you have one that’s 100% clean.

You’re definitely on the right track though. Way to go mang:rasta:


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OnlineLogicaL ChaosM
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Re: Cloning Tek, best way to multiply? [Re: Actionjunkie]
    #26449820 - 01/24/20 11:45 AM (4 years, 5 days ago)

Yes, Cloning narrows down the genetics while a spore print is basically random genetics.

Best way to clone is to clone a shroom on one plate, let it colonize then place slices of mycelium on other clean plates. Then transfer to grain or an LC.


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OfflineA.k.aM
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Re: Cloning Tek, best way to multiply? [Re: LogicaL Chaos]
    #26449834 - 01/24/20 11:49 AM (4 years, 5 days ago)

Yeah you’re right about prints being random, I meant the post above talking about saving lc or whatever to not have to pay for spores again.


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LAGM2020


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OfflineHuskies
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Re: Cloning Tek, best way to multiply? [Re: A.k.a]
    #26450141 - 01/24/20 02:23 PM (4 years, 5 days ago)

You get one good flush and you'll never need to buy spores again breh


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InvisibleGoombas69
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Registered: 12/15/19
Posts: 15
Re: Cloning Tek, best way to multiply? [Re: feldman114]
    #26450156 - 01/24/20 02:40 PM (4 years, 5 days ago)

Quote:

feldman114 said:
Full stop.
Peroxide can’t be used to clean anything. Also, how would you “clean” an LC? With agar, the term refers to making transfers of transfers from transfers, until you’re sure you have one that’s 100% clean.

You’re definitely on the right track though. Way to go mang:rasta:




Thanks feldman114!
I had read that hydrogen peroxide can be a last ditch effort to salvage mycelium in a LC since mycelium can survive hydrogen peroxide and Trich cannot. I now read that you cannot get enough hydrogen peroxide to kill Trich mold without killing the mycelium too, so thanks for setting me straight. I'll stick with agar transfers if I get into trouble. I always make prints after my first flush of a new mushroom so hopefully I never have to deal with this problem.

Thanks again!


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Offlinefeldman114
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Re: Cloning Tek, best way to multiply? [Re: Goombas69]
    #26450159 - 01/24/20 02:43 PM (4 years, 5 days ago)

Quote:

Goombas69 said:
Quote:

feldman114 said:
Full stop.
Peroxide can’t be used to clean anything. Also, how would you “clean” an LC? With agar, the term refers to making transfers of transfers from transfers, until you’re sure you have one that’s 100% clean.

You’re definitely on the right track though. Way to go mang:rasta:




Thanks feldman114!
I had read that hydrogen peroxide can be a last ditch effort to salvage mycelium in a LC since mycelium can survive hydrogen peroxide and Trich cannot. I now read that you cannot get enough hydrogen peroxide to kill Trich mold without killing the mycelium too, so thanks for setting me straight. I'll stick with agar transfers if I get into trouble. I always make prints after my first flush of a new mushroom so hopefully I never have to deal with this problem.

Thanks again!




Using agar? Doing your own research?? Utilizing the search function?????!

Breaking all the noob stereotypes - good on ya, bud.
You’re on your way to mush cult greatness:derdance:


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OfflineeLeSDenes
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Re: Cloning Tek, best way to multiply? [Re: LogicaL Chaos]
    #26450170 - 01/24/20 02:53 PM (4 years, 5 days ago)

Quote:

LogicaL Chaos said:
Yes, Cloning narrows down the genetics while a spore print is basically random genetics.

Best way to clone is to clone a shroom on one plate, let it colonize then place slices of mycelium on other clean plates. Then transfer to grain or an LC.



Quote:



:whathesaid:

Look how nice they they are straight from the pin


some of them a few transfers away:




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