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Offlineflattoptony
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Questions regarding cloning
    #7330296 - 08/24/07 05:46 PM (13 years, 6 months ago)

1) Would making an LC of a sport print be a more efficient way of cloning by sticking a syringe through the stem?

2) Also, why is it better to stick the syringe through the stem?

3) Would it be possible to isolate a single spore?

4) Is a petri dish LC faster than in jars, how does this really work?


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OfflineNibin
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Re: Questions regarding cloning [Re: flattoptony]
    #7330340 - 08/24/07 05:58 PM (13 years, 6 months ago)

1) If it involves spores it isn't cloning

2) Dunno

3) A single spore wouldn't work. To get shrooms you need two spores to combine their DNA. That is how fungi sexually reproduce

4)Petri dish LC? You mean Agar techniques? You have a solid, nutritious surface for the mycelia to grow on, and by forming different colonies you can isolate the stronger strains and use them to grow with


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Offlinesimplystoned
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Re: Questions regarding cloning [Re: flattoptony]
    #7330420 - 08/24/07 06:27 PM (13 years, 6 months ago)

Quote:

flattoptony said:
1) Would making an LC of a sport print be a more efficient way of cloning by sticking a syringe through the stem?

2) Also, why is it better to stick the syringe through the stem?

3) Would it be possible to isolate a single spore?

4) Is a petri dish LC faster than in jars, how does this really work?




I think perhaps you're misusing some of the terms here, bud. I'll try to answer the questions best I can.

1)Making a LC with a spore print is not cloning, it is simply innoc'ing an lc and making more viable mushroom solution. An LC made by a spore print will be "multispore", meaning there will be multiple genetics each time you inoculate.

2) By sticking a syringe into the middle of a mushroom stem you are obtaining a small piece of mushroom tissue which can be used to help you isolate the strain. If this piece of tissue were put onto agar and then used to inoculate an LC then you would have a liquid culture of isolated genetics.

3) Isolate a single spore? uh...

4) A petri dish liquid culture? There's no such thing. Do some more reading and brush up on your terms.


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


"Your pain is the breaking of the shell which encloses your understanding. It is the bitter potion by which the physician within you heals your sick self. Therefore trust the physician, and drink his remedy in silence and tranquility:

For his hand, though heavy and hard, is guided
by the tender hand of the Unseen,
And the cup he brings, though it burn your lips,
has been fashioned of the clay which the Potter
has moistened with His own sacred tears." - Kahlil Gibran


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InvisibleBacchus
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Re: Questions regarding cloning [Re: simplystoned]
    #7330461 - 08/24/07 06:42 PM (13 years, 6 months ago)

Quote:

4) A petri dish liquid culture? There's no such thing. Do some
more reading and brush up on your terms.




Not so fast there... Why can't one agitate a colonized petri dish with sterile water and collect the resulting mycelium-rich solution?


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Invisiblejeetered
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Re: Questions regarding cloning [Re: flattoptony]
    #7330464 - 08/24/07 06:42 PM (13 years, 6 months ago)

Quote:

flattoptony said:
1) Would making an LC of a sport print be a more efficient way of cloning by sticking a syringe through the stem?

2) Also, why is it better to stick the syringe through the stem?

3) Would it be possible to isolate a single spore?

4) Is a petri dish LC faster than in jars, how does this really work?




1) core sample cloning is a great method, making a "Sport Print SYringe" is still a multispore LC.. not a clone..

2) I like the stipe for cloning, as it's meatier, the cap is full of basidia, and the flesh is flimsy.. therefore, you run the risk of no flesh in your "clone sample" and also introducing basidium to your clone, (spores) which would negate the act of cloning.

3)Diluting a syringe, down to 5% spores, helps in isolation, isolating spores to a single spore, would require microscopic work, and using agar/petri.. are you prepared for that?

4) Petri dishes are not LC's, they are used for cloning and isolating myc that shows the most rhizomorphic traits, (tomentose mycelia doesn't like to fruit as well)

once you've isolated on agar the clone/isolate you desire, moving it to a master grain jar, or an LC is simple..

keep in mind, all clones, isolates degrade genetically over time, and then you will have to start at square one again..

it is recommended that one make stock slants, to store, and maintain, a clone... having 10 slants of the same clone, keeps the genetics alive and kicking, keep it dormant, then a few years later, pop out a slant, incubate it, watch for growth, put in another master grain jar.. (for g2g clone transfers)

using mineral oil atop your clone slant, or petri, allows the myc to go dormant and not contam..

read more in the forums and archives, you'll figure it out.

i suggest getting "the magic mushroom growers guide"
or watch youtube, or download the .pdf's

Quote:

Nibin said:
1) If it involves spores it isn't cloning

2) Dunno

3) A single spore wouldn't work. To get shrooms you need two spores to combine their DNA. That is how fungi sexually reproduce


so, you can possibly germinate two spores, get good clamp connections, and form diakaryosis after TIME, yes, TIME< and shaking your grains!!!

but, i don't think you are at that level yet AT ALL...

peace nugkas.


4)Petri dish LC? You mean Agar techniques? You have a solid, nutritious surface for the mycelia to grow on, and by forming different colonies you can isolate the stronger strains and use them to grow with




1.*this is correct..

2....lollers if you don't know about cloning why are you answering?

3. ***This is partially correct, but two spores may mate to be monokaryotic, which wont fruit in our sense, diakaryiotic is whatyou are looking for, which, is 2 mated pairs... (four sexes)
mushrooms have four sexes, not two.....

4. yes there is petri TO LC.. not petri LC... but petri cutting TO lc... or, g2g to a master grain jar, then g2g from there...



Quote:

Bacchus said:
Quote:

4) A petri dish liquid culture? There's no such thing. Do some
more reading and brush up on your terms.





Not so fast there... Why can't one agitate a colonized petri dish with sterile water and collect the resulting mycelium-rich solution?







one can take cuttins from a petri, and make an LC, yes, that's a pretty standard procedure..

using a water wash, would be a waste of a good dish...



Edited by jeetered (08/24/07 07:17 PM)


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