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InvisibleBuddha420
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Registered: 05/15/10
Posts: 411
Loc: Mobil, Alabama
Re: Cloning vs print culturing [Re: PrimalSoup]
    #13391432 - 10/26/10 05:35 PM (13 years, 5 months ago)

Quote:

PrimalSoup said:

This is what's going senescent on you now?  No wonder. :shrug:  Again, you don't propagate from clone to clone to clone - you want to make an isolate (clone) that performs well and preserve it at the earliest point in time you can.  Then you return to that preserved culture for new grows.

Peace,
-PS




i have not been tracking my genetic progress for years...untill recent...mostly everything i make though, performs well.


how do you identify a "monoculture"

im nore sure i fully understand what it means really.


--------------------
Psychedelic Salon - changing our minds, one thought at a time

Religion - George Carlin, R.I.P.

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Re: Cloning vs print culturing [Re: Buddha420]
    #13391559 - 10/26/10 06:04 PM (13 years, 5 months ago)

Quote:

Buddha420 said:
how do you identify a "monoculture"




a monoculture on agar will show only a single sector of growth on agar...

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InvisibleBuddha420
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Re: Cloning vs print culturing [Re: total]
    #13391995 - 10/26/10 07:41 PM (13 years, 5 months ago)

Quote:

total said:
Quote:

Buddha420 said:
how do you identify a "monoculture"




a monoculture on agar will show only a single sector of growth on agar...





Yeah, I've seen posts...and my myc. doesnt sector at all on agar. When I make dish transfers, I always take a tiny fiber. I use a magnifying glass, and a very fine scalpel to peel as tiny of a fiber as I can. Previously I made petri dish transfers by cutting a triangle out of the agar.


Here are my petris right now:






What is the difference between transfering a very small fiber and cutting out a large triangle?


Also...what exactly is a monoculture..I understand that it doesn't sector on agar, but is there a genetic analogy in mammals for mono vs multi-spore cultures?

Edited by Buddha420 (10/26/10 07:53 PM)

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Re: Cloning vs print culturing [Re: Buddha420]
    #13392350 - 10/26/10 08:49 PM (13 years, 5 months ago)

I have yet to start with my agar journey...
But looking back at all iv read, it looks like you have multiple sectors on CLMS on both A and B...
Hold them upto a strong light and look through the back...you will see dividing lines...
Maybe post pics, i swear i see them from this distance :shrug:

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InvisibleBuddha420
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Re: Cloning vs print culturing [Re: total]
    #13393268 - 10/27/10 12:03 AM (13 years, 5 months ago)

Quote:

total said:
I have yet to start with my agar journey...
But looking back at all iv read, it looks like you have multiple sectors on CLMS on both A and B...
Hold them upto a strong light and look through the back...you will see dividing lines...
Maybe post pics, i swear i see them from this distance :shrug:





you're right... i do see sectoring...

that clone was taken from a non-cloned fruit...so those 2 dishes are multi-spore cultures? what should i do? pick the sector with the most rhizomorphic myc. and transfer that??

also..if you're trying to isolate a sub-strain, so you transfer from a single sector, after a few transfers the myc. will degenerate. So you are pretty much forced to spawn it, and only then continue the strain isolation??


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Re: Cloning vs print culturing [Re: Buddha420]
    #13393374 - 10/27/10 12:20 AM (13 years, 5 months ago)

Quote:

Buddha420 said:
that clone was taken from a non-cloned fruit...so those 2 dishes are multi-spore cultures? what should i do? pick the sector with the most rhizomorphic myc. and transfer that??

also..if you're trying to isolate a sub-strain, so you transfer from a single sector, after a few transfers the myc. will degenerate. So you are pretty much forced to spawn it, and only then continue the strain isolation??




A. You should take a small piece from each sector and xfer to another plate and see if it grows a monoculture...once you have your various monocultures you should fruit each out to see which is best for you...you can then keep that plate as a master plate(M1), make 5 more plates from it(production plates we will call P1-5), store the master, these production plates p1-5 should last you quite awhile...just take a small piece from one of the p1-5 and place on another plate, grow it out, and then use wedges from these plates to inoculate your grain jars...

B. Id say you have more xfers that what you might think you do...

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InvisibleBuddha420
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Re: Cloning vs print culturing [Re: total]
    #13393584 - 10/27/10 01:41 AM (13 years, 5 months ago)

Quote:

total said:
Quote:

Buddha420 said:
that clone was taken from a non-cloned fruit...so those 2 dishes are multi-spore cultures? what should i do? pick the sector with the most rhizomorphic myc. and transfer that??

also..if you're trying to isolate a sub-strain, so you transfer from a single sector, after a few transfers the myc. will degenerate. So you are pretty much forced to spawn it, and only then continue the strain isolation??




A. You should take a small piece from each sector and xfer to another plate and see if it grows a monoculture...once you have your various monocultures you should fruit each out to see which is best for you...you can then keep that plate as a master plate(M1), make 5 more plates from it(production plates we will call P1-5), store the master, these production plates p1-5 should last you quite awhile...just take a small piece from one of the p1-5 and place on another plate, grow it out, and then use wedges from these plates to inoculate your grain jars...





As you can see, I have my own cataloging system. I just pick a name such as CLMS (clone monster shroom) the 1 after S is generation of that genetic line. And then I label each dish A-D or B, and add the letter to the name with each transfer...so the amount of letters in the name alows me to see how many transfers have been made, so CLMS1A abd CLMS1B would be the 2 initial "mothers".


then anything with more than 1 letter are production plates...reason I like the lettering is because I always work with petris in that order, A,B,C,D...so if something happens during the procedure...I make a mistake...I can always note that specific dish.


Quote:



B. Id say you have more xfers that what you might think you do...





can you elaborate? not sure what you mean here.





also...im not equipped to store live myc. I dont have the test tubes, nor a dedicated fridge. I cannot really "save" the mother plates...i can only track the outcome and save a print.


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Psychedelic Salon - changing our minds, one thought at a time

Religion - George Carlin, R.I.P.

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OfflinePrimalSoup
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Re: Cloning vs print culturing [Re: Buddha420]
    #13395487 - 10/27/10 02:08 PM (13 years, 5 months ago)

Quote:

I dont have the test tubes, nor a dedicated fridge.




Test tubes are cheap on eBay and elsewhere.   

Not any fridge at all - or one that you can't control?  That would be a drawback.  There are other ways to store master cultures, from what I've read (dessication, under mineral oil) but I've not used them. :shrug:

Quote:

also..if you're trying to isolate a sub-strain, so you transfer from a single sector, after a few transfers the myc. will degenerate.




What "few transfers"?  It's less transfers than what you seem to be doing now in propagation - you just transfer from the "best" sector until it stops sectoring - short growth like that doesn't make it senescent.  It's continued growth without stepping back to earlier stored cultures that weakens the strain, from what I've seen anyway...

Quote:

Also...what exactly is a monoculture..I understand that it doesn't sector on agar, but is there a genetic analogy in mammals for mono vs multi-spore cultures?




A monoculture is one expressing the same genetics thoughout - mono culture. 

Most humans are monocultures in many respects, but some humans are mosaics (like blue and green eyes), sometimes caused by a failed twining - and recent research has shown mosaic traits in humans going generations back into ancestry, in certain organs at least (like blood). But then things are rarely as simple as they might seem. :lol:

A better comparison would be a lab mouse, bred over many generations to a genotype and expressing predictable traits.  Or, say, dog breeds.  Or almost any long isolated species.

From what I can see you're doing great work, I wouldn't worry about any of this overmuch.  I like the looks of your phenotypical fruits too!

Peace
-Ps

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InvisibleBuddha420
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Posts: 411
Loc: Mobil, Alabama
Re: Cloning vs print culturing [Re: PrimalSoup]
    #13396162 - 10/27/10 04:25 PM (13 years, 5 months ago)

thanks, Primal.

Here's the deal...whenever I started the culture from a live tissue, it always sectors...then with the 1st xfer, sectoring usually drops...but its always been like that, I am yet to produce #1 dish (the one with the actual live tissue) that doesn't sector...

Even when I isolate that single sector myc. and then spawn it, then take a clone off the fruit...it still sectors at in the original dish.


--------------------
Psychedelic Salon - changing our minds, one thought at a time

Religion - George Carlin, R.I.P.

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Re: Cloning vs print culturing [Re: Buddha420]
    #13396720 - 10/27/10 06:33 PM (13 years, 5 months ago)

Im just gonna hit on the points primal hit on, as i just got out of surgery and still abit loopy in the reading
Quote:

I dont have the test tubes, nor a dedicated fridge.




Get some PP Culture tubes on Cynmar...Its like $4/25
The only reason you need tubes is for long-term storage...
Plates should store in the fridge for atleast 6mnths, at which point you transfer abit off the old plate onto a new plate...that should give you another 6mnths :smile:

As far as not having a fridge...im sure you have a fridge that you can wrap plates in parafilm, throw them in a ziplock, and then throw them in a brown paper bag...You dont have to label your plates "p. cubensis"
As far as storing your tubes once you get them, you cover your slants with sterile distilled water, and freeze them...Iv been told this will preserve the tubes for 20+ years.

Quote:

also..if you're trying to isolate a sub-strain, so you transfer from a single sector, after a few transfers the myc. will degenerate.




You should only be making 3-4 transfers deep off your master culture...once master culture could last you forever if they would last that long :wink:
I have a few questions of my own as well...Maybe some older hands can help and answer.
Is it ALL about transfers that degrade it? or is there a time factor involved as well?
Starting with MS, what should be my MAX number of petri transfers?

Quote:

Also...what exactly is a monoculture..I understand that it doesn't sector on agar, but is there a genetic analogy in mammals for mono vs multi-spore cultures?




I used the sectoring because its was the easiest way i could put it... :shrug:
I really dont know the technical details, Id say its a single STRAIN of the mycelium, but some might confuse that with these names we put on the cubes...

I just really like participating in Adv Mycology...things move slower here...
I think im gonna go eat some pain meds and put my ankle up...

total

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