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Manna Inspired
NOT-SO-NOOB
Registered: 11/28/07
Posts: 67
Loc: Under the stars
Last seen: 16 years, 3 months
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clone vs. going all the way and isolating your clone
#7710241 - 12/03/07 10:43 AM (16 years, 3 months ago) |
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It has been my experience that a clone on agar will continue to sector. And after several transfer isolations of rhizo's, I will get an isolate from the clone that no longer sectors itself. Then into the run it out and see how well it works for you stage ( this in itself a good reason to isolate several rhizo sectors from the clone, something is bound to come out bountiful). My question is; I see some teks where people are talking about cloning and then just using the resulting myc. growth as there clone (taking tissue and regrowing it in a substrate). Is this adequate in it's purpose in having same resulting final product ( as in my clone isolates)?
-------------------- EXPEIRINCE IS IN THE KNOWLEDGE AND NOT THE RATING.
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veda_sticks
Cultivator
Registered: 07/29/07
Posts: 14,191
Loc: UK
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Re: clone vs. going all the way and isolating your clone [Re: Manna Inspired]
#7710510 - 12/03/07 11:56 AM (16 years, 3 months ago) |
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interesting question, as I thought that clonning a shroom would result in 1 sub strain of myc.
I'm not that clued up on the growth of myc and sectoring etc. someone with some experience will hopefully stop by.
-------------------- PF TEK - writeup by EvilMushroom666 Lets Grow Mushrooms - RogerRabbit & RoadKills website with sample videos plus the full PF TEK video series. Alot of great information - BUY THE DVD Cakes can and will pin! - So you think cakes suck for pins. Your wrong Franks Simple Coir/Verm Tek Franks Proper Pasturisation Tek Franks Spawning To Bulk - Monotub Professor Pinheads RTV Injection Port Tek Foo Mans No Soak WBS Prep Tek
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Manna Inspired
NOT-SO-NOOB
Registered: 11/28/07
Posts: 67
Loc: Under the stars
Last seen: 16 years, 3 months
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Re: clone vs. going all the way and isolating your clone [Re: veda_sticks]
#7710515 - 12/03/07 11:58 AM (16 years, 3 months ago) |
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Hopefully. I get the feeling most people talking about cloning stop there. I have really consistent results w/ going the further step, just wondering if those that stop short of isolation get consistent results as I have.
-------------------- EXPEIRINCE IS IN THE KNOWLEDGE AND NOT THE RATING.
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Premedman1
Assistant to the insistent
Registered: 12/21/05
Posts: 2,376
Loc: South of Sanity
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Re: clone vs. going all the way and isolating your clone [Re: Manna Inspired]
#7710575 - 12/03/07 12:07 PM (16 years, 3 months ago) |
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I think to answer your question, you first must answer why a single tissue sample will sector on agar when all of its genetic makeup is, in theory, identical (genetic drift notwithstanding).
-------------------- Build a man a fire, he is warm for the night. Set a man on fire, he is warm for the rest of his life.
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Wronguy
Registered: 03/05/05
Posts: 4,450
Last seen: 12 years, 8 months
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Re: clone vs. going all the way and isolating your clone [Re: Manna Inspired]
#7710582 - 12/03/07 12:08 PM (16 years, 3 months ago) |
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Quote:
Manna Inspired said: My question is; I see some teks where people are talking about cloning and then just using the resulting myc. growth as there clone (taking tissue and regrowing it in a substrate). Is this adequate in it's purpose in having same resulting final product ( as in my clone isolates)?
Short answer is no. Isolating a strain from a clone ensures a greater chance you are isolating a strain that will actually produce fruits. Now in the scheme of things, speaking in general terms, you have a better chance of selecting a strain that will produce fruits during isolation if you're taking from rhizomorphic sectors when compared to tomentose, but this does not always happen.
I have always isolated sectors using cloned mushroom tissue grown out on agar and have never cultivated a non-fruiting strain. No matter how you slice it, you are never going to know if you've selected the best strain unless you isolate every rhizomorphic sector from agar and try them all. I'm speaking of spores though, not mushroom tissue. This is the nature of the beast unfortunately.
Does that answer your question?
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Manna Inspired
NOT-SO-NOOB
Registered: 11/28/07
Posts: 67
Loc: Under the stars
Last seen: 16 years, 3 months
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Re: clone vs. going all the way and isolating your clone [Re: Wronguy]
#7710608 - 12/03/07 12:13 PM (16 years, 3 months ago) |
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yes. As I knew the answer just needed reenforcement.
think to answer your question, you first must answer why a single tissue sample will sector on agar when all of its genetic makeup is, in theory, identical (genetic drift notwithstanding).
It has been stated many times that it is believed that a fruit has not one but many sub-strains that make up the form(so to myself this is answered).
-------------------- EXPEIRINCE IS IN THE KNOWLEDGE AND NOT THE RATING.
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Manna Inspired
NOT-SO-NOOB
Registered: 11/28/07
Posts: 67
Loc: Under the stars
Last seen: 16 years, 3 months
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Re: clone vs. going all the way and isolating your clone [Re: Manna Inspired]
#7710614 - 12/03/07 12:14 PM (16 years, 3 months ago) |
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Thanks
-------------------- EXPEIRINCE IS IN THE KNOWLEDGE AND NOT THE RATING.
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Wronguy
Registered: 03/05/05
Posts: 4,450
Last seen: 12 years, 8 months
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Re: clone vs. going all the way and isolating your clone [Re: Premedman1]
#7710619 - 12/03/07 12:14 PM (16 years, 3 months ago) |
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Quote:
Premedman1 said: I think to answer your question, you first must answer why a single tissue sample will sector on agar when all of its genetic makeup is, in theory, identical (genetic drift notwithstanding).
Sectoring from a clone grown on agar is not only possible, it happens more often than not in my experience. It is not uncommon to have a fruit comprised of more than one strain. When it does happen, you have to try every sector to get the best results from the genetic makeup of that clone.
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Manna Inspired
NOT-SO-NOOB
Registered: 11/28/07
Posts: 67
Loc: Under the stars
Last seen: 16 years, 3 months
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Re: clone vs. going all the way and isolating your clone [Re: Wronguy]
#7710636 - 12/03/07 12:18 PM (16 years, 3 months ago) |
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Right on. This has been my experience.
-------------------- EXPEIRINCE IS IN THE KNOWLEDGE AND NOT THE RATING.
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Premedman1
Assistant to the insistent
Registered: 12/21/05
Posts: 2,376
Loc: South of Sanity
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Re: clone vs. going all the way and isolating your clone [Re: Wronguy]
#7710679 - 12/03/07 12:22 PM (16 years, 3 months ago) |
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Quote:
Wronguy said:
Quote:
Premedman1 said: I think to answer your question, you first must answer why a single tissue sample will sector on agar when all of its genetic makeup is, in theory, identical (genetic drift notwithstanding).
Sectoring from a clone grown on agar is not only possible, it happens more often than not in my experience. It is not uncommon to have a fruit comprised of more than one strain. When it does happen, you have to try every sector to get the best results from the genetic makeup of that clone.
I see. Thanks. It's still hard for me to wrap my mind around one fruit containing more than one set of genetics, but I guess that's the nature of the beast.
-------------------- Build a man a fire, he is warm for the night. Set a man on fire, he is warm for the rest of his life.
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Wronguy
Registered: 03/05/05
Posts: 4,450
Last seen: 12 years, 8 months
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Re: clone vs. going all the way and isolating your clone [Re: Premedman1]
#7710801 - 12/03/07 12:42 PM (16 years, 3 months ago) |
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Cloning allows you to preserve the genetic makeup of the fruit, not isolate a particular strain contained within. The problem with cloning is that it could, depending on the genetic signature of the fruitbody, contain multiple strains. If your goal is to create an exact copy of that clone, you will, in theory, have to allow all the containing strains to colonize and fruit.
Now comes the hard part. How do we prove that all the strains embedded within that fruitbody will perform in the exact same manner as its original? Unfortunately, I don't think we can. All we can do is inoculate the substrate with the cloned tissue, whether grown on grain spawn or liquid media, and allow the mycelium to do what it does. I don't think the mycelia are pre-programmed to grow in the exact same manner as its predecessor, only contain the same genetic signature.
This topic appears to be getting in to discussion that may be beyond my expertise, so I'll stop it there.
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Manna Inspired
NOT-SO-NOOB
Registered: 11/28/07
Posts: 67
Loc: Under the stars
Last seen: 16 years, 3 months
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Re: clone vs. going all the way and isolating your clone [Re: Wronguy]
#7711868 - 12/03/07 05:00 PM (16 years, 3 months ago) |
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I think we are done here. I am.Thanks
-------------------- EXPEIRINCE IS IN THE KNOWLEDGE AND NOT THE RATING.
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veda_sticks
Cultivator
Registered: 07/29/07
Posts: 14,191
Loc: UK
Last seen: 4 years, 2 months
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Re: clone vs. going all the way and isolating your clone [Re: Manna Inspired]
#7712198 - 12/03/07 06:03 PM (16 years, 3 months ago) |
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cue RR to come in and drop in a nugget of information thats not been said yet
-------------------- PF TEK - writeup by EvilMushroom666 Lets Grow Mushrooms - RogerRabbit & RoadKills website with sample videos plus the full PF TEK video series. Alot of great information - BUY THE DVD Cakes can and will pin! - So you think cakes suck for pins. Your wrong Franks Simple Coir/Verm Tek Franks Proper Pasturisation Tek Franks Spawning To Bulk - Monotub Professor Pinheads RTV Injection Port Tek Foo Mans No Soak WBS Prep Tek
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Pr0_X
CultivationLifer
Registered: 01/18/06
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Re: clone vs. going all the way and isolating your clone [Re: veda_sticks]
#7745892 - 12/11/07 04:55 PM (16 years, 3 months ago) |
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Bump..
I'm currently interested in this same subject, I took tissue samples from one mushroom and placed them on agar as well as tissue from a 2nd mushroom from the same flush, Would it be possible to isolate out the most rhizomorph section of growth if I continued to transfer sections to there six generation and then plucked out the best growth from there?
Would this be a waste of my time or would I actually be getting the best genetics the mycelium offers?
-------------------- It's okay to hurt my feelings cause you know, they're so numb anyway. but I guess it's what I get for being to fuckin stupid to stay away - Jake - Support the FSR at www.fsrcanada.com and www.fsre.nl
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tahoe
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Re: clone vs. going all the way and isolating your clone [Re: Pr0_X]
#7745928 - 12/11/07 05:00 PM (16 years, 3 months ago) |
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explain the difference between isolating and sectoring. How can you tell when an isolate stops sectoring? Does it just grow 1 rhizo straight across the agar? How many transfers away from a spore laden petri do you have to be before it becomes an isolate?
-------------------- Stop experimenting half way through your first grow. Grow it to maturity, watch it, learn from it. Do this a few times then experiment with different ideas and figure out what works best for you. My Legacy https://www.shroomery.org/forums/showflat.php/Number/22140987#22140987 Teh=The I need to proofread
Edited by tahoe (12/11/07 05:00 PM)
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Wronguy
Registered: 03/05/05
Posts: 4,450
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Re: clone vs. going all the way and isolating your clone [Re: Pr0_X]
#7746792 - 12/11/07 08:09 PM (16 years, 3 months ago) |
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Quote:
Pr0_X said: Would it be possible to isolate out the most rhizomorph section of growth if I continued to transfer sections to there six generation and then plucked out the best growth from there?
Absolutely, but keep in mind that you would be isolating the best strain from the genetic makeup of the clone. You also need to do several transfers and literally try them all to find out which one is the best performer. It is a lengthy process, but only has to be done once if you take the time to grow out each sector.
Quote:
Pr0_X said: Would this be a waste of my time or would I actually be getting the best genetics the mycelium offers?
Again, you're only going to be able to isolate the strains contained within the genetic makeup of the selected clone. If you're using the term "mycelium" to suggest the entire substrate as a whole, the answer would be a resounding no. You would need to isolate from spores and could easily spend months isolating sectors and months again growing each sector out.
Quote:
tahoe said: explain the difference between isolating and sectoring. How can you tell when an isolate stops sectoring? Does it just grow 1 rhizo straight across the agar?
Isolating is the actual task of sector selection by which each sector will be transfered to another petri dish, and another, and another, until you have narrowed it down to a single strain that no longer sectors out.
Sectoring can be defined as unique independent growth coming from a single inoculation area.
Quote:
tahoe said: How many transfers away from a spore laden petri do you have to be before it becomes an isolate?
25, 50, 100? There is not set answer to this unfortunately. It all depends on the amount of spores that germinate. I think RogerRabbit may have a better answer for this one.
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