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OfflineHallucinogenist
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Re: LC Cloning vs Isolating [Re: J3illy]
    #9374171 - 12/05/08 05:09 PM (15 years, 3 months ago)

Just checking....  ^_^

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OfflineNibin
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Re: LC Cloning vs Isolating [Re: Hallucinogenist]
    #9374611 - 12/05/08 06:22 PM (15 years, 3 months ago)

He is not the only "guru" here but he certainly is one of the most active ones so it is easier to find posts by him.

Here are some of the things that make us trust his advice. There are a few other users who also share many of these characteristics

He has been growing edibles and actives for many many years and as a few other select members actually takes his studies quite scientifically.

He knows Stamets personally and actually disagrees with him on some things, which shows he doesn't just take whatever he says as absolute truth, but backs things up with his own experience.

He has grown shrooms on the most random things like a Bra, a bible, weed, etc.

He has contributed things like the shotgun terrarium, the videos etc.

There are a few other old hands which are just as knowledgeable as him, but they aren't as prolific when posting in the normal mush cult forum.


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Newcomers guide-----> For all things shroomy

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Offlinealkylbenzene23
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Re: LC Cloning vs Isolating [Re: Nibin]
    #9374764 - 12/05/08 06:48 PM (15 years, 3 months ago)

A... Bra?
:orly:
:lol:


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Invisiblearp180
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Re: LC Cloning vs Isolating [Re: alkylbenzene23]
    #9374819 - 12/05/08 06:54 PM (15 years, 3 months ago)

check out RR's Gallery and you will see it.


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OfflineHallucinogenist
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Re: LC Cloning vs Isolating [Re: arp180]
    #9374833 - 12/05/08 06:57 PM (15 years, 3 months ago)

Wow...  Well, I'm officially a believer.  haha

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OfflineLightShedder
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Re: LC Cloning vs Isolating [Re: Hallucinogenist]
    #9374884 - 12/05/08 07:02 PM (15 years, 3 months ago)

Not only have I done LC cloning before and got varied results from the LC, but I have personally, on multiple occasions in fact, seen a piece of flesh (even from LC clones) sector out into as many as 6 substrains on agar.


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OfflineHallucinogenist
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Re: LC Cloning vs Isolating [Re: LightShedder]
    #9374916 - 12/05/08 07:06 PM (15 years, 3 months ago)

Okay, a question now...


LC is obviuosly something that even an amateur shoomerite can do (myself for example).  Is agar just as simple, or does it actually require more skill and scientific knowledge/equipment?

Also, how exactly is isolation done?

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OfflineStimpy913
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Re: LC Cloning vs Isolating [Re: alkylbenzene23]
    #9374962 - 12/05/08 07:12 PM (15 years, 3 months ago)

Quote:

alkylbenzene23 said:
A... Bra?
:orly:
:lol:




And a burrito.  I like his videos. If you try to read up on things here, most of the time people blow it way out of proportion with complication and uncessesary things. In RR's video its' just straight facts, and a good demonstration of idiotproof methods.  If i never saw the video, and just read the shroomery, there'd be no growing because the videos show you how easy it really is.  Plus it's cool to see someone grow oysters & shiitake off cakes. I never knew that could be done before.


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InvisibleJ3illy
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Registered: 10/18/08
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Re: LC Cloning vs Isolating [Re: Hallucinogenist]
    #9375082 - 12/05/08 07:29 PM (15 years, 3 months ago)

Quote:

Hallucinogenist said:
LC is obviuosly something that even an amateur shoomerite can do (myself for example).  Is agar just as simple, or does it actually require more skill and scientific knowledge/equipment?

Also, how exactly is isolation done?




From reading about both, I'd say working w/ agar is gonna be a step up from just making an LC.  Sterility has to be totally on point w/ agar, just cuz that shit is used specifically for growing out bacteria and spores - so any spec of contamination that may be present will flourish.  You need sterility in all aspects obviously, but w/ agar it's MAJOR.

And isolation is getting spores/flesh on agar, and once it starts growing, there will be separate sub-strains.  You then transfer a small piece of a sub-strain to a new agar dish, and once that's the ONLY one growing - you've isolated it.

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OfflineHallucinogenist
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Re: LC Cloning vs Isolating [Re: J3illy]
    #9375144 - 12/05/08 07:37 PM (15 years, 3 months ago)

And how do you identify the sub-strain on the agar?  Microscopically, or am I misunderstanding the Agar process?

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InvisibleJ3illy
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Re: LC Cloning vs Isolating [Re: Hallucinogenist]
    #9375192 - 12/05/08 07:45 PM (15 years, 3 months ago)

A single substrain is like it's own totally differentiated growth.  For example in this pic, the top 1 is isolated, but the bottom 1 looks like many sub-strains growing together.  You'd transfer a small piece from the bottom 1, and wait to see how it grows, and keep transferring until it's isolated.



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OfflineHallucinogenist
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Re: LC Cloning vs Isolating [Re: J3illy]
    #9375203 - 12/05/08 07:47 PM (15 years, 3 months ago)

You mean you'd actually FRUIT the piece from the bottom right?  To see if it's correct?

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OfflineiMtRiPpIn
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Re: LC Cloning vs Isolating [Re: Hallucinogenist]
    #9375242 - 12/05/08 07:53 PM (15 years, 3 months ago)

Quote:

Hallucinogenist said:
You mean you'd actually FRUIT the piece from the bottom right?  To see if it's correct?




I think he means you would take a piece from the bottom picture, and put it on agar and grow it out, and keep repeating the process until you get growth that looks like the top picture.


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OfflineLightShedder
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Re: LC Cloning vs Isolating [Re: Hallucinogenist]
    #9375255 - 12/05/08 07:55 PM (15 years, 3 months ago)

I use the grocery store agar tek. It is just as easy as LC. I use short half-pint jars (or maybe they are pint I am not sure). The lids are upside down (for easy opening) closed tightly with a 1/4x1/4 inch hole in the middle stuffed with polyfill.

In the grocery store agar tek he calls for "1/2 of a 9g strip" of solid agar. I like to find the powder agar from health stores and just weigh out 4.5 grams. It is easier then breaking up all the strips.

Also make sure if you get the powder agar that it doesn't have suger or something mixed in. You want plain agar agar.

Once you have an agar plate with one substrain on it (like picture 1 above) then you can make multiple agar plates from it to have a good amount of it stored up. Then you can take a tiny chunk of the mycelium from the plate and drop it in a LC. You can turn the LC into 100s more LCs and then have a huge amount of this isolate stored for use. Just make sure to test out the strain before you go about making so much culture so you know if you even want it:cool:.


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InvisibleJ3illy
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Re: LC Cloning vs Isolating [Re: iMtRiPpIn]
    #9375292 - 12/05/08 08:03 PM (15 years, 3 months ago)

Quote:

iMtRiPpIn said:
Quote:

Hallucinogenist said:
You mean you'd actually FRUIT the piece from the bottom right?  To see if it's correct?




I think he means you would take a piece from the bottom picture, and put it on agar and grow it out, and keep repeating the process until you get growth that looks like the top picture.




Exactly.  You cut a really small piece of it, and transfer to a new plate.  Then once you have the isolated plate, it's like your master copy - all you gotta do is cut a little piece and you can start a new plate, use it for an LC or inoculation.  You'd just keep it in the fridge and it'd be good for a while.  For really long-term storage, they have these things called "slants", and the isolated culture will last years.

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InvisibleJ3illy
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Re: LC Cloning vs Isolating [Re: LightShedder]
    #9375312 - 12/05/08 08:07 PM (15 years, 3 months ago)

Quote:

LightShedder said:
I use the grocery store agar tek. It is just as easy as LC. I use short half-pint jars (or maybe they are pint I am not sure). The lids are upside down (for easy opening) closed tightly with a 1/4x1/4 inch hole in the middle stuffed with polyfill.




How does everything work out, do you get many contams?  I definitely wanna start working w/ agar, but I'm just kinda discouraged about thinking of contams.  If I had a flow hood it'd be a different story, lol.  Do you do everything in a glove-box?

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Invisiblearp180
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Re: LC Cloning vs Isolating [Re: J3illy]
    #9375319 - 12/05/08 08:08 PM (15 years, 3 months ago)

"From reading about both"

I am glad to see you started your post with this.  I am just starting to read about agar and isolation work.  The description and pictures you cited were very helpful.  Thanks J3illy

Edited by arp180 (12/05/08 08:10 PM)

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OfflineiMtRiPpIn
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Re: LC Cloning vs Isolating [Re: arp180]
    #9375349 - 12/05/08 08:13 PM (15 years, 3 months ago)

Quote:

arp180 said:
"From reading about both"

I am glad to see you started your post with this.  I am just starting to read about agar and isolation work.  The description and pictures you cited were very helpful.  Thanks J3illy




yeah thanks J3illy the pictures were defintiely worth a thousand words.  my head starts spinning when i hear terms like "sectoring" and shit but seeing the beginning and end result spoke volumes.


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Invisiblenw_shroomy
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Re: LC Cloning vs Isolating [Re: arp180]
    #9375366 - 12/05/08 08:15 PM (15 years, 3 months ago)

I'm sure this can be answered with search but since its being discussed.
How long after transferring to agar does it take to grow before its time to transfer to the next agar?


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OfflineLightShedder
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Re: LC Cloning vs Isolating [Re: J3illy]
    #9375367 - 12/05/08 08:16 PM (15 years, 3 months ago)

Yeah I do everything in a sealed still-air glovebox. The only way to pretty much ensure 0% contaminations everytime IMO.


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