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goodman1
Stranger
Registered: 01/14/18
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cloning vs strain isolation
#24914918 - 01/15/18 02:42 AM (7 years, 3 days ago) |
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Is there any significant difference between cloning tissue vs isolating a strain after multi spore inoculation of agar?
Gearing up for my first grow, and I am a bit of a science nerd, although biology isn't my field. I was wondering if I should try to isolate a single strain with agar, or just do a multispore grow and clone the best the looking fruit.
I will be doing a monotub by the way.
any advice/input appreciated, please correct me wherever my logic is wrong as im a noob, thank you.
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Letsgrowstones
Pebble



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Re: cloning vs strain isolation [Re: goodman1]
#24914925 - 01/15/18 02:55 AM (7 years, 3 days ago) |
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Hi,
I am a noob too,
They are the same thing how ever there is a post on here where someone has gone through 27+ agar to agar transfers in search on an iso.
Cloning from tissue ensures you have an iso...
My suggestion is do a multi to agar, this will help diminish the chance of contamination drastically. If contam then agar health to another agar.
Then move clean and healthy ms mycelium to grain to mono.
From here clone 8 good mushrooms on agar, do 2 agars per clone(1to keep for later grows, other to grain/monotub)
Keep the best toss the rest.
Regards.
Ps. Skilled growers please correct this post how you see fit
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goodman1
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if I took some tissue from inside the stem I'd think that would minimize contamination. Working on building a sort of custom ghetto flowhood type system that I think will work quite well.
27+ agar transfers is the reason why im thinking cloning tissue is best. Also this way I can dehydrate it and sample it for potency right after I clone.
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nube424

Registered: 12/03/17
Posts: 6,063
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Re: cloning vs strain isolation [Re: goodman1]
#24914947 - 01/15/18 03:27 AM (7 years, 3 days ago) |
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Use a SAB. Works just as well as a flow hood. And toom up agar sectoring. Thats strain isolation. Its only an iso if theres no sectors in the mycellium. Clone is the best way to start this too. And yes myc from the inside of the stem butt. Just tear the stem open lengthwise and take a biopsy
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SadTurkey
ayy


Registered: 10/21/17
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Re: cloning vs strain isolation [Re: nube424] 1
#24914955 - 01/15/18 03:34 AM (7 years, 3 days ago) |
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Another advantage of cloning is that you can select a strain that grows and pins readily.
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nube424

Registered: 12/03/17
Posts: 6,063
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Re: cloning vs strain isolation [Re: SadTurkey]
#24914957 - 01/15/18 03:39 AM (7 years, 3 days ago) |
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Quote:
SadTurkey said: Another advantage of cloning is that you can select a strain that grows and pins readily.
Yea take clones from ur best performing substrate and look for a big fruit in a cluster.
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goodman1
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Re: cloning vs strain isolation [Re: nube424]
#24914961 - 01/15/18 03:45 AM (7 years, 3 days ago) |
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ive heard that more then one strain is active when you clone from a multispore fruit, because its basically a combo of a few different strains into one fruit body.
So im not actually sure if cloning gives an legit iso.
may be best to isolate from agar rather then clone, then its just hit or miss, get lucky.
Edited by goodman1 (01/15/18 03:45 AM)
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nube424

Registered: 12/03/17
Posts: 6,063
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Re: cloning vs strain isolation [Re: goodman1]
#24914964 - 01/15/18 03:48 AM (7 years, 3 days ago) |
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Quote:
goodman1 said: ive heard that more then one strain is active when you clone from a multispore fruit, because its basically a combo of a few different strains into one fruit body.
So im not actually sure if cloning gives an legit iso.
may be best to isolate from agar rather then clone, then its just hit or miss, get lucky.
Well yea once u clone itll have multiple strains. U have to transfer each sector to new agar plates till u stop seeing sectoring. THEN its an isolate.
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pacmanbreed


Registered: 10/12/16
Posts: 3,829
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Re: cloning vs strain isolation [Re: nube424]
#24914994 - 01/15/18 04:34 AM (7 years, 2 days ago) |
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Not all isolate are good though. Better to innoc grains after you clean a clone to agar keeping some of the strains.
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Threads from God89
Pioneer.......


Registered: 08/10/17
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Re: cloning vs strain isolation [Re: pacmanbreed]
#24915161 - 01/15/18 07:45 AM (7 years, 2 days ago) |
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Developing a true monoculture (isolate) can take forever. Cloning is a great way to preserve and obtain certain genetics.
I was told by hamloaf that is is best to wait to select your clone specimen until the fruit has matured and the veil begins to tear.... HOWEVER do not wait until it drops spores, that is TOO LATE. You want it to be mature so that you can observe the fruits proper characteristics, but not dropping spores because once it drops spores the fruit begins to die...... bodhissata and hamloaf both agreed on this and that is info I obtained from those two knowledgable Cultivators.
hamloaf told me to remove the cap from the stem when preparing to clone...... And take an inner tissue sample from the inside of the stipe, from underneath where the cap was attached. Be ready to do your work and transfer the cloning fruit directly to your SAB or work area after harvesting! 
Maybe this will be helpful....? 
https://www.shroomery.org/forums/showflat.php/Number/24909129#24909129
-------------------- Admirer of Bushido....... Seeker of Truth, and Knowledge.
I Will Keep the Fire going, and the Good Wolf fed.......
( We Are All Threads from God.)
Edited by Threads from God89 (01/15/18 07:48 AM)
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Crombie
Cheese Boat Captain



Registered: 11/17/17
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Quote:
Letsgrowstones said:
Cloning from tissue ensures you have an iso...
Incorrect. The number of strains will likely be significantly reduced compared to MS, but there's no guarantee a mushroom will contain a single strain.
Otherwise, you've got a good plan. I intend to go this route once my tub is finished. I don't have the patience to do 50 agar transfers from MS
-------------------- Sailing the seas of cheese!
My beginner journal https://www.shroomery.org/forums/showflat.php/Number/24845768
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RomeoPapa
Jackass of All Trades



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Re: cloning vs strain isolation [Re: Crombie]
#24916367 - 01/15/18 05:13 PM (7 years, 2 days ago) |
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Being new myself I just like to focus on CLEAN spawn. It's sometimes easier said than done. Absolutely start with agar. Don't worry yourself with isolates or clones until you have a few successful grows under your belt and get a feel for things.
-------------------- It's better to have it and not need it
Than it is to need it and not have it.
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cronicr



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Re: cloning vs strain isolation [Re: RomeoPapa]
#24916454 - 01/15/18 05:45 PM (7 years, 2 days ago) |
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Either way you have lots of testing to do. I like to start my spores..get a clean culture and start a grow but also keep transferring and also leave plates to pin and clone those...each isolate and each clone will need testing. Advantage of isolates from spores..younger cell lines less division. Advantage of clibes...proven fruiting and ya already have an idea of what you are working with.
Many say isolates from spores is not ideal as you can end up with a non fruiting or shitty fruiting strain but fact is most varieties are pretty dam stable and odds are low you end up with either.
-------------------- 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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NotJonHamm
Stranger Danger

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Re: cloning vs strain isolation [Re: cronicr]
#24916707 - 01/15/18 07:08 PM (7 years, 2 days ago) |
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I think it would be more worthwhile to clone good performing fruits, at first anyways. An experienced grower, PussyFart, did an experiment **link** where he used 10 PE isolates from good PE fruits and grew them out with widely varying results with only 6/10 tubs producing fruit. I think the lesson to be learned was to not find an isolate for the sake of finding an isolate (especially if you're going from MS straight to agar - you'll have no idea what you're isolating), but to find an isolate once you know you have a good set of genes that produce quality fruits.
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mushpunx
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Re: cloning vs strain isolation [Re: cronicr]
#24917634 - 01/16/18 07:18 AM (7 years, 1 day ago) |
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Quote:
cronicr said: Either way you have lots of testing to do. I like to start my spores..get a clean culture and start a grow but also keep transferring and also leave plates to pin and clone those...each isolate and each clone will need testing. Advantage of isolates from spores..younger cell lines less division. Advantage of clibes...proven fruiting and ya already have an idea of what you are working with.
Many say isolates from spores is not ideal as you can end up with a non fruiting or shitty fruiting strain but fact is most varieties are pretty dam stable and odds are low you end up with either.
 Right on Cron.
I love agar work so I do strain isolation as a side project. Takes a lot of testing, I like to use substrate bottles fruited in monotub to test lots of cultures side by side at once under the same condition. Its not as tedious as people make it out to be.
But for a starting cultivator like you OP, I'd get good at growing and cloning first
--------------------

Amateur Mycologists United
AMU Q&A
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mentat
Thinker



Registered: 04/24/15
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Loc: Arrakis
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Re: cloning vs strain isolation [Re: mushpunx]
#24917677 - 01/16/18 07:55 AM (7 years, 1 day ago) |
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very good, informative read thanks everyone... 
there is one thing that escapes me in cloning and isolation topic.
let's follow cron's method above.. spores started on agar, then transferred and many clean cultures are developing. Some are maturing, thus A2G is done, some left to pin happily in the dish. A2G master spawn jar, may or may not fruit, or even different sections of 1 healthy dish would surely give different results/yields since it is a clean MS culture afterall..
suppose at this point, we take the first winner pin from one of those happily pinning plates. put it in another dish, let it grow and observe the speed, texture. at this point, as far as i understand it still doesn't bring monoculture, but the gene pool is narrowed down quite a bit.
at this point two things I fail to grasp: why not cloning a healthy pin from agar plate would give guaranteed fruiting strains? although several (maybe even 10) strains running in the shroom, wouldn't it be a fairly good chance to obtain a fruit-sure culture. why we should still test it on grains?
and secondly, suppose we did test the pin-culture on grains. and it was a success, fruits emerging from sub after conditions introduced.. then what? all from the beginning, spores on agar, then expanded and pinned, then cloned and expanded into grains and fruited again... to avoid mutations and senescence, don't we have to cease cloning over again and take spores at some stage and start over?
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bodhisatta 
Smurf real estate agent


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Re: cloning vs strain isolation [Re: mentat]
#24917870 - 01/16/18 09:41 AM (7 years, 1 day ago) |
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If you clone a pin of course you have a guarantee to have a fruiting culture.
I've never seen a non fruiting culture anyway.
I wouldn't worry about senescence right now. Most people don't run into those kinds of problems. You're really going to grow only the one culture for a few years? Even still if you had good storage techniques it wouldn't even matter. Like how we have strains of edibles that have been around dozens of years.
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Bird_Guts
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Re: cloning vs strain isolation [Re: bodhisatta]
#24917883 - 01/16/18 09:45 AM (7 years, 1 day ago) |
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Cloning is the way to go imo
Professor pinhead had an insane PE isolate, but it took him like 3 years to get it
If you want a true monoculture then I would take a clone to agar and start isolating from there
--BG--
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mentat
Thinker



Registered: 04/24/15
Posts: 126
Loc: Arrakis
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Re: cloning vs strain isolation [Re: bodhisatta]
#24917891 - 01/16/18 09:48 AM (7 years, 1 day ago) |
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yes, thanks bod..
i also came across with pussyfart's reply somewhere 4 years ago, very illuminating.
Quote:
PussyFart said: Well from spores to fruit is generation 1.
If you clone that fruit, grow it out and fruit it again, this would be generation 2.
You will hit the end of the road very soon doing it this way.
But if you keep going back to that first set of cloned genetics, you can make a ton of transfers and keep it going.
Think about it, RR has had cultures for 15-20 years, how?
Every years he takes his slants and transfers it to a dish, grows it out, and knocks up a new master slant.
finally i get the dynamics of it.
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cronicr



Registered: 08/07/11
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Re: cloning vs strain isolation [Re: bodhisatta]
#24918262 - 01/16/18 01:07 PM (7 years, 1 day ago) |
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Quote:
bodhisatta said: If you clone a pin of course you have a guarantee to have a fruiting culture.
I've never seen a non fruiting culture anyway.
I wouldn't worry about senescence right now. Most people don't run into those kinds of problems. You're really going to grow only the one culture for a few years? Even still if you had good storage techniques it wouldn't even matter. Like how we have strains of edibles that have been around dozens of years.
Idk bod...some states pushing for legalizing get those isolates ready Tbh my two best cultures are crs and cambodian but neither is a clone or an isolate...simple ms cultures that just produce
-------------------- 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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