|
Some of these posts are very old and might contain outdated information. You may wish to search for newer posts instead.
|
Tookitooki
Mycological Fabricator



Registered: 07/28/16
Posts: 1,157
Loc: Nowhere
|
How far do you stretch your culture?
#23974593 - 01/01/17 11:19 PM (7 years, 28 days ago) |
|
|
You start with a Ms grow, you put some clones to agar. clean them up and get them back to grain and test them. You find a really good culture. So you put a slant together for safe keeping and you work with what's left of the agar plate.
Weather you go LC, grain masters, or a pile of agar plates from the original agar plates.
How far can you take that culture before it starts giving you issues?
And is one method preferred over another to help preserve that culture?
I've read that you don't want to clone a clone. But if you keep spawning out the mycilium, at what point does it start to degrade.
|
Intelligentxfruit
Earth Hippy


Registered: 01/06/13
Posts: 1,545
|
Re: How far do you stretch your culture? [Re: Tookitooki]
#23974654 - 01/01/17 11:58 PM (7 years, 28 days ago) |
|
|
How far you expand depends on the media you're going through. agar/lc can be expanded much more than g2g. thats why batches of master jars are made to expand as much with grain from the start. I think 4-5 g2g is the accepted number? I do 3
Slants are the prefer for long term storage.
You can clone a clone, but once you reach the 3rd+ generation it degrades pretty rapidly.
|
wildernessjunkie
Reshitivest


Registered: 06/13/10
Posts: 8,118
Loc: HTTP 404 Not Found
|
Re: How far do you stretch your culture? [Re: Tookitooki]
#23974662 - 01/02/17 12:01 AM (7 years, 28 days ago) |
|
|
You shouldn't need to clone clones unless you've somehow lost the master culture. If you maintain that culture properly then it has the potential to last decades.
If you learn to do agar, you can keep very young genetics for a long period of time. And if you can do LI, then having a stack of petris in the fridge can stretch a hell of a long way.
With that said, I remember seeing RR post that he didnt start running into senescence issues until he reached his 8th G2G transfers.
|
Thedillestpickle
cultured



Registered: 02/02/16
Posts: 1,170
Loc: Canada
Last seen: 3 years, 7 months
|
|
What if you clone from a very mature fruit after it has opened veil and sporulated? Would that clone have scenescence already?
I'm trying to decide if I need to ditch this culture I cloned or if it's just my inoculation method that needs work.
Edited by Thedillestpickle (01/02/17 12:18 AM)
|
wildernessjunkie
Reshitivest


Registered: 06/13/10
Posts: 8,118
Loc: HTTP 404 Not Found
|
|
Whats wrong with your cloned culture?
Some of the best commercial cultures come from clones. The average hobby grower will never run into problems with senescence. Take this math into account, and try to visualize this as it plays out. Gimme a bit of leeway since Ive been on the wine for a couple hours and might fuck this all up....
You clone to agar. You do three transfers to be sure its clean and then inoculate a single jar of grain. You then take that jar and use it to G2G at a 1:10 ratio.
1:10=10 jars
Each of those is spawned 1:10 as well, which is reasonable, and we follow what RR said about 8 generations.
10x10=100
10x100=1000
10x1000=10000
10x10000=100000....
And thats probably far enough. Thats more spawn than anyone I know, including a commercial operation can deal with. If you can visualize how much space that would take to store, and we arent even to the 8 generations of spawn yet. Some additional space would be required for substrate on top of all that too. The amount of mycelium at that point is far, far greater than a hobby grower is likely to deal with.
Whats going on with the culture you have? Why are you considering ditching it?
|
Thedillestpickle
cultured



Registered: 02/02/16
Posts: 1,170
Loc: Canada
Last seen: 3 years, 7 months
|
|
Sorry maybe I'm hijacking the OP's thread with my question.
I'm not the OP, I just have my own issue and I'm trying to figure out if it's scenescence.
The issue is that I really like the fruits it grows but it grows very slowly, sometimes stalling out and often goes bacterial and also very tomentose growth.
|
tump
ban the undead



Registered: 03/17/16
Posts: 2,383
Last seen: 6 years, 10 months
|
|
Honestly after gene 3 i always screwed up the tranfers. The myc does age and start to weaken in any state. Even still based on this logic you can turn one clean plate of ms into many things but easy count lets say you make one quarts jars of oats and one pint of lc form little wedge. Best results is to let lc and oats colonized. Tranfer lc when done two ten more qts lc. And do g2g twice for 100 good jars. This is 2 months into the process. Make 20 monos and clone five fruit on agar form each mono. That is 100 fruit clones iso. Make one pints lc out of each iso clean plates. Knock up 20 jars form each pint of master lc to test the culture in fruiting. If you like the culture you can make ten more qts of lc which will make 100 grain jars easy and fast colonized. Once the lc is done eating all food in liquid it needs to be put in cold storage. Same with master plates. If you ever need more lc ms then go back to master ms jar. Even with never getting past gene 3 with the lc you can grow 1000's of pounds of mushrooms. Mushrooms that buy spawn form big spawn productions know that the culture they are growing is much older then most of them . example is oyster strain 3815 i believe is 60+ years old. You can expland with proper control of culture more then any mushroom kingpins need in life time
|
Tookitooki
Mycological Fabricator



Registered: 07/28/16
Posts: 1,157
Loc: Nowhere
|
|
not looking for mass spawn. but the idea of keeping a good culture for a long time. maybe i should of worded the subject a little differently.
Quote:
wildernessjunkie said: If you learn to do agar, you can keep very young genetics for a long period of time. And if you can do LI, then having a stack of petris in the fridge can stretch a hell of a long way.
With that said, I remember seeing RR post that he didnt start running into senescence issues until he reached his 8th G2G transfers.
so let say G2G (from agar wedge) has a limit of 8 generations, does the same apply for generations of agar?
so you take a slant, bring it to agar (call it P1). once it grows out you make 10 more plates. if you took a piece of the agar from P1, and put it back to a slant, would that be gen 2?
before i get flamed, i'm not looking for no kingpin business, I just have questions in my head that i couldn't locate a satisfying answer for.
Its not like its hard to throw a MS grow together and snag another clone. Im just trying to get a better understanding of the growth and aging process of mycelium. Life span before vigor loss. with so much information on here, and no matter how much research and sorting you do, sometimes you start seeing what looks like contradicting info. Since the first time i got involved in mycology and this site (reader only back in 2008 for about a year and a half) so much has evolved and changed. last time around i was after the fruits, this time around its for the passion i developed previous. i just wish i had more time to devote to this hobby
|
van hatton
Still a noob



Registered: 11/23/14
Posts: 5,617
Loc: Michigan
|
Re: How far do you stretch your culture? [Re: Tookitooki]
#23975303 - 01/02/17 07:56 AM (7 years, 28 days ago) |
|
|
-------------------- If I ever give out misinformation please inform me so I can have the correct information. Tmethyl said: Chuck Norris once roundhouse kicked a monotub that wasn't pinning fast enough. The force of the kick rearranged the genetics of the mushrooms, we now call them Penis Envy. Caps McGee said:
Fun part is figuring out what works best for you
|
amidogen
see you on the other side

Registered: 05/07/16
Posts: 1,782
|
Re: How far do you stretch your culture? [Re: van hatton]
#23975406 - 01/02/17 08:53 AM (7 years, 28 days ago) |
|
|
-------------------- The biggest trip of my life was realizing all of the events and actions described in posts made by this account were never real and had never actually happened, but were instead the delusional ramblings of a severely mentally ill human being. I just had to move on for my own good. I love you all.
|
wildernessjunkie
Reshitivest


Registered: 06/13/10
Posts: 8,118
Loc: HTTP 404 Not Found
|
Re: How far do you stretch your culture? [Re: amidogen]
#23975683 - 01/02/17 10:48 AM (7 years, 28 days ago) |
|
|
The P Scalle is something that Stamets came up with. And yes it has some value.
As you make transfers, technically yes the culture is expanding and getting older cell lines. However, we go back to the practical application of this idea. Think of the volume of a jar of spawn, and compare that to the volume in a petri dish.
If we take fully colonized petri dishes and use them to fill up a spawn jar, how many would it take? I dont know, but quite a few right? At the end, you would still end up with about the same amount of cell expansion.
Ive seen master cultures that are sold with P Values over 100. These are still fairly young genetics. And when you are dealing with clones, theres no real way to determine what the initial P value would be. Theres not a lot of practical application for this for the average hobbiest.
Edit: If you are getting slow tomentose growth. Try switching to a low nutrient agar. I bet your myc gets real aggressive as it gets hungry.
Edited by wildernessjunkie (01/02/17 10:50 AM)
|
Tookitooki
Mycological Fabricator



Registered: 07/28/16
Posts: 1,157
Loc: Nowhere
|
|
i don't have any issues or anything. it was just a answer that i tried finding but wasn't happy with what i found.
so your saying that since agar requires considerably less growth then a grain master, that the gen limit of agar would be significantly more then the "8 gen limit" seen by RR on G2G. It makes total sense.
so if you pull your slant out once a year to do some work, and put a piece of that first agar plate back in to a new slant, you could continue that cycle longer then most practical applications with out any real issues.
:Take this as a hypothetical, non practical situation, informational purpose only:
you get a very unique culture. the potency hits, the fruits are amazing size and unique looks, its a fast growing culture. you throw the clone on a slant, but try to stabilize the genetics thru its prints, but it never shows its face again. but you now have a culture you hope to maintain until you walk away from the hobby.
as long as you were smart about things, your telling me that the culture has the potential to last a long time to come? i fully understand nothing is a guarantee.
sometimes certain questions come to mind and i just have to get a good elaborate answer to satisfy my brain.
|
bodhisatta 
Smurf real estate agent


Registered: 04/30/13
Posts: 61,889
Loc: Milky way
|
Re: How far do you stretch your culture? [Re: Tookitooki]
#23976105 - 01/02/17 01:00 PM (7 years, 28 days ago) |
|
|
go until performance drops off usually 6-12 months depending on how you expand. much longer if you go back to refrigerated masters.
most of these answers you got look like people trying to reiterate things they read somewhere as if they know like the back of their hand how it works
|
enlightenment
alchemist


Registered: 08/09/09
Posts: 1,647
Loc: Europe
Last seen: 8 months, 21 days
|
Re: How far do you stretch your culture? [Re: bodhisatta]
#23976116 - 01/02/17 01:05 PM (7 years, 28 days ago) |
|
|
100%
|
wildernessjunkie
Reshitivest


Registered: 06/13/10
Posts: 8,118
Loc: HTTP 404 Not Found
|
Re: How far do you stretch your culture? [Re: enlightenment]
#23976363 - 01/02/17 02:27 PM (7 years, 28 days ago) |
|
|
Quote:
as long as you were smart about things, your telling me that the culture has the potential to last a long time to come?
If its slanted and maintained, yes. I used the same master slants for 5 years before I moved to a new state.
|
bodhisatta 
Smurf real estate agent


Registered: 04/30/13
Posts: 61,889
Loc: Milky way
|
|
a good example is edible and medicinal cultures. Lentinula Edodes 75 "shiitake 75" is a strain which has been around at least 10 years (can't find out when it was brought about)
|
Tookitooki
Mycological Fabricator



Registered: 07/28/16
Posts: 1,157
Loc: Nowhere
|
Re: How far do you stretch your culture? [Re: bodhisatta]
#23977309 - 01/02/17 08:57 PM (7 years, 28 days ago) |
|
|
That's awesome to know. Thanks for being patient with me.
|
|