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KauaiOrca
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Registered: 08/12/08
Posts: 3,131
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CLONE WATER Experiment Updates ...
#23916317 - 12/11/16 01:24 PM (7 years, 1 month ago) |
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Given that mushroom stems are generally fairly hollow, I had an idea in terms of cloning that would be an easy, one-step process. I would take a sterile syringe with about 2-3 ML's of water and then cut the mushroom near the base with a red hot knife, At the base, the stem is not hollow so through the solid, meaty bottom, the red hot needle goes up and into hollow part of the stem, always staying inside the stem ... I then inject the sterile water into the hollow stem, leaving the needle in there ... shake it a bit an then suck the water back into the needle, which never left the stem ... take the needle out, heat it quickly and then inject the water into a small jar of RGS.
I tried the poke the stem with the needle and try to get a piece of it technique that never worked for me, but am wondering if anyone's done this? Theoretically, it's sterile in the hollow part of the mushroom, right? I'd think the water injected would pick up enough cells to get a small jar of RGS going which could then be slurried for liquid innoculant. Could also, I guess do it to agar too. Any thoughts?
-------------------- "The universe is endless, limitless and infinite. Any effort to define it's boundaries is an attempt to overcome ignorance. We are physical, mental and spiritual beings ... there is no beginning and there is no end. There is only memory. Our repeated loss of memory experiences create the illusion of beginnings and ends. Immortality is the ability to retain full memory through all consciousness transformations. Loss of memory is man's greatest curse and, in very real terms, death." -- Ancient Taoist Master
Edited by KauaiOrca (01/22/17 08:43 AM)
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mushboy
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Re: Anyone Tried Cloning This Way??? [Re: KauaiOrca]
#23916321 - 12/11/16 01:28 PM (7 years, 1 month ago) |
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i guess you could do that but it seems like too much trouble. i have shaky hands. ill end up poking through the stem and sticking myself.
you can use any piece of the mushroom. its recommenced to use the inside of the stipe because you can get a cleaner piece of tissue.
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tump
ban the undead



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Re: Anyone Tried Cloning This Way??? [Re: mushboy]
#23916371 - 12/11/16 01:46 PM (7 years, 1 month ago) |
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Agar it. Or lc it like that tek says
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KauaiOrca
Waterman


Registered: 08/12/08
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Re: Anyone Tried Cloning This Way??? [Re: mushboy]
#23916376 - 12/11/16 01:47 PM (7 years, 1 month ago) |
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I have pretty steady hands ... It's actually really easy if you pick a stout, straight mushroom. The reason I want to try this is I can just use the water I sucked back to inoculate a small jar of grass seed and should hopefully avoid any step of cleaning on agar, etc.
Anyway I'm trying, will report back results but was just curious if anyone else had tried this because hopefully, can get enough cells without even opening the mushroom up at all.
-------------------- "The universe is endless, limitless and infinite. Any effort to define it's boundaries is an attempt to overcome ignorance. We are physical, mental and spiritual beings ... there is no beginning and there is no end. There is only memory. Our repeated loss of memory experiences create the illusion of beginnings and ends. Immortality is the ability to retain full memory through all consciousness transformations. Loss of memory is man's greatest curse and, in very real terms, death." -- Ancient Taoist Master
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KauaiOrca
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Registered: 08/12/08
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Re: Anyone Tried Cloning This Way??? [Re: tump]
#23916389 - 12/11/16 01:51 PM (7 years, 1 month ago) |
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Quote:
tump said: Agar it. Or lc it like that tek says
How would it pick up any contams? The stem would never have even been exposed to open air. The bottom of the stem (meaty part) would be cut with a red hot blade ... If needle and water is sterile, there is no way for contams to get in unless the inside of the stem is contaminated which seems very unlikely, right?
-------------------- "The universe is endless, limitless and infinite. Any effort to define it's boundaries is an attempt to overcome ignorance. We are physical, mental and spiritual beings ... there is no beginning and there is no end. There is only memory. Our repeated loss of memory experiences create the illusion of beginnings and ends. Immortality is the ability to retain full memory through all consciousness transformations. Loss of memory is man's greatest curse and, in very real terms, death." -- Ancient Taoist Master
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spirit_shadow
Feature not a bug



Registered: 08/15/11
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Re: Anyone Tried Cloning This Way??? [Re: KauaiOrca]
#23916460 - 12/11/16 02:12 PM (7 years, 1 month ago) |
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You could skip the knife and just heat the needle red hot then poke it, let it cool, then squirt the water.....im going to try this sometime thanks for the idea lol
-------------------- ERROR 418 IM A TEAPOT.....(this account is automated, all posts related to illegal activities or advice thereof are strictly from numerous online sites and are for informational purposes only)- Circa 2011 Ban lotto
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Pastywhyte
Say hello to my little friend



Registered: 09/15/12
Posts: 37,810
Loc: Canada
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In the end you are going to have a weak culture, that takes forever to recover, from a mature specimen, in a format not conducive to long term storage. Obvious contam vectors aside (how do you sterilize the outside of the fruit? Answer is you can't, best you can do is sanitize) this might seem appealing to someone with extremely limited experience but, it's going to lose its appeal when the results are in.
Also, what if the fruit isn't hollow?
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cronicr



Registered: 08/07/11
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Re: Anyone Tried Cloning This Way??? [Re: Pastywhyte]
#23916503 - 12/11/16 02:27 PM (7 years, 1 month ago) |
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The inside is not necessarily sterile..if you ever cloned an outdoor fruit you would know this
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  It doesn't matter what i think of you...all that matters is clean spawn I'm tired do me a favor
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spirit_shadow
Feature not a bug



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Re: Anyone Tried Cloning This Way??? [Re: Pastywhyte]
#23916505 - 12/11/16 02:27 PM (7 years, 1 month ago) |
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Quote:
Pastywhyte said: In the end you are going to have a weak culture, that takes forever to recover, from a mature specimen, in a format not conducive to long term storage. Obvious contam vectors aside (how do you sterilize the outside of the fruit? Answer is you can't, best you can do is sanitize) this might seem appealing to someone with extremely limited experience but, it's going to lose its appeal when the results are in.
Also, what if the fruit isn't hollow?

... touche'
-------------------- ERROR 418 IM A TEAPOT.....(this account is automated, all posts related to illegal activities or advice thereof are strictly from numerous online sites and are for informational purposes only)- Circa 2011 Ban lotto
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KauaiOrca
Waterman


Registered: 08/12/08
Posts: 3,131
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Re: Anyone Tried Cloning This Way??? [Re: Pastywhyte]
#23916561 - 12/11/16 02:48 PM (7 years, 1 month ago) |
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Quote:
Pastywhyte said: In the end you are going to have a weak culture, that takes forever to recover, from a mature specimen, in a format not conducive to long term storage. Obvious contam vectors aside (how do you sterilize the outside of the fruit? Answer is you can't, best you can do is sanitize) this might seem appealing to someone with extremely limited experience but, it's going to lose its appeal when the results are in.
Also, what if the fruit isn't hollow?

All good points. I was thinking of using a fruit that's about half grown so its younger and it's easy to tell if it's hollow by just squeezing it a bit. You may be right on the weak culture aspect, though ... hadn't thought of that. I'll see how it compares to snagging pins from a cluster (how I do it now) and going to Agar. Main thought was I could possibly skip 2-3 agar transfers ... Go straight to RGS like Violet's method.
-------------------- "The universe is endless, limitless and infinite. Any effort to define it's boundaries is an attempt to overcome ignorance. We are physical, mental and spiritual beings ... there is no beginning and there is no end. There is only memory. Our repeated loss of memory experiences create the illusion of beginnings and ends. Immortality is the ability to retain full memory through all consciousness transformations. Loss of memory is man's greatest curse and, in very real terms, death." -- Ancient Taoist Master
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cronicr



Registered: 08/07/11
Posts: 61,436
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Re: Anyone Tried Cloning This Way??? [Re: KauaiOrca] 1
#23916566 - 12/11/16 02:49 PM (7 years, 1 month ago) |
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Why try cut corners...we take from the base and the cap for good reason.
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  It doesn't matter what i think of you...all that matters is clean spawn I'm tired do me a favor
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KauaiOrca
Waterman


Registered: 08/12/08
Posts: 3,131
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Re: Anyone Tried Cloning This Way??? [Re: cronicr]
#23916605 - 12/11/16 03:05 PM (7 years, 1 month ago) |
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Quote:
cronicr said: Why try cut corners...we take from the base and the cap for good reason.
Wasn't really aware this is cutting corners. I generally use pins to start new cultures and have had good success with it ... Growing them really clean invitro saves steps too ... never really tried cloning a mature fruit as it seemed like a PITA ...
-------------------- "The universe is endless, limitless and infinite. Any effort to define it's boundaries is an attempt to overcome ignorance. We are physical, mental and spiritual beings ... there is no beginning and there is no end. There is only memory. Our repeated loss of memory experiences create the illusion of beginnings and ends. Immortality is the ability to retain full memory through all consciousness transformations. Loss of memory is man's greatest curse and, in very real terms, death." -- Ancient Taoist Master
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cronicr



Registered: 08/07/11
Posts: 61,436
Loc: Van Isle
Last seen: 2 years, 9 days
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Re: Anyone Tried Cloning This Way??? [Re: KauaiOrca]
#23916616 - 12/11/16 03:08 PM (7 years, 1 month ago) |
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Im talking about the op's idea of squirting water into fruits
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  It doesn't matter what i think of you...all that matters is clean spawn I'm tired do me a favor
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Pastywhyte
Say hello to my little friend



Registered: 09/15/12
Posts: 37,810
Loc: Canada
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Re: Anyone Tried Cloning This Way??? [Re: KauaiOrca]
#23916651 - 12/11/16 03:20 PM (7 years, 1 month ago) |
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Quote:
KauaiOrca said:
Quote:
Pastywhyte said: In the end you are going to have a weak culture, that takes forever to recover, from a mature specimen, in a format not conducive to long term storage. Obvious contam vectors aside (how do you sterilize the outside of the fruit? Answer is you can't, best you can do is sanitize) this might seem appealing to someone with extremely limited experience but, it's going to lose its appeal when the results are in.
Also, what if the fruit isn't hollow?

All good points. I was thinking of using a fruit that's about half grown so its younger and it's easy to tell if it's hollow by just squeezing it a bit. You may be right on the weak culture aspect, though ... hadn't thought of that. I'll see how it compares to snagging pins from a cluster (how I do it now) and going to Agar. Main thought was I could possibly skip 2-3 agar transfers ... Go straight to RGS like Violet's method.
You will see your success rate drop and clean a lot of mold.
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KauaiOrca
Waterman


Registered: 08/12/08
Posts: 3,131
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Re: Anyone Tried Cloning This Way??? [Re: Pastywhyte]
#23916733 - 12/11/16 03:48 PM (7 years, 1 month ago) |
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Quote:
Pastywhyte said:
Quote:
KauaiOrca said:
Quote:
Pastywhyte said: In the end you are going to have a weak culture, that takes forever to recover, from a mature specimen, in a format not conducive to long term storage. Obvious contam vectors aside (how do you sterilize the outside of the fruit? Answer is you can't, best you can do is sanitize) this might seem appealing to someone with extremely limited experience but, it's going to lose its appeal when the results are in.
Also, what if the fruit isn't hollow?

All good points. I was thinking of using a fruit that's about half grown so its younger and it's easy to tell if it's hollow by just squeezing it a bit. You may be right on the weak culture aspect, though ... hadn't thought of that. I'll see how it compares to snagging pins from a cluster (how I do it now) and going to Agar. Main thought was I could possibly skip 2-3 agar transfers ... Go straight to RGS like Violet's method.
You will see your success rate drop and clean a lot of mold.
How does mold enter the equation? Is there mold inside a half grown fruit? If I cut the stem with a red hot knife and use a red hot needle to enter into the stem, where does mold get a foothold? Not trying to be difficult, I just honestly can't see how mold would get inside the needle. I also live in the high desert where mold levels, I think are a lot lower.
-------------------- "The universe is endless, limitless and infinite. Any effort to define it's boundaries is an attempt to overcome ignorance. We are physical, mental and spiritual beings ... there is no beginning and there is no end. There is only memory. Our repeated loss of memory experiences create the illusion of beginnings and ends. Immortality is the ability to retain full memory through all consciousness transformations. Loss of memory is man's greatest curse and, in very real terms, death." -- Ancient Taoist Master
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Kenetic
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Re: Anyone Tried Cloning This Way??? [Re: KauaiOrca]
#23916757 - 12/11/16 03:58 PM (7 years, 1 month ago) |
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For starters it wouldn't grow very fast if you went directly to rgs. You can get a enough myc on agar in a few days to accelerate your spawn by at least a week. Plus pretty much guarantee it's clean.
-------------------- Todo Cambia    DMT said: Everyone know's me, they just don't know it yet
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KauaiOrca
Waterman


Registered: 08/12/08
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Re: Anyone Tried Cloning This Way??? [Re: Kenetic]
#23916769 - 12/11/16 04:02 PM (7 years, 1 month ago) |
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Quote:
kenetic said: For starters it wouldn't grow very fast if you went directly to rgs. You can get a enough myc on agar in a few days to accelerate your spawn by at least a week. Plus pretty much guarantee it's clean.
It would grow faster than spores, wouldn't you think? Maybe I'm wrong here.
-------------------- "The universe is endless, limitless and infinite. Any effort to define it's boundaries is an attempt to overcome ignorance. We are physical, mental and spiritual beings ... there is no beginning and there is no end. There is only memory. Our repeated loss of memory experiences create the illusion of beginnings and ends. Immortality is the ability to retain full memory through all consciousness transformations. Loss of memory is man's greatest curse and, in very real terms, death." -- Ancient Taoist Master
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Peteyboy
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Re: Anyone Tried Cloning This Way??? [Re: Kenetic]
#23916771 - 12/11/16 04:03 PM (7 years, 1 month ago) |
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Random cloning question but relevant to OPS post..is it a bad idea to clone a mature specimen just beginning to tear it's veil on the first flush or should only pins be used for cloning?
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cronicr



Registered: 08/07/11
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Re: Anyone Tried Cloning This Way??? [Re: KauaiOrca]
#23916774 - 12/11/16 04:04 PM (7 years, 1 month ago) |
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It would be faster to clone properly on agar..pins are great to clone but a more mature fruit will work fine as well
--------------------
  It doesn't matter what i think of you...all that matters is clean spawn I'm tired do me a favor
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Peteyboy
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Re: Anyone Tried Cloning This Way??? [Re: cronicr]
#23916788 - 12/11/16 04:08 PM (7 years, 1 month ago) |
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Awesome thanks Cron...You da man!
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