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OfflineTHK107
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Keeping isolated strains healthy
    #23197339 - 05/06/16 11:01 PM (8 years, 8 months ago)

So I've been doing a lot of research on cloning and isolating with agar.  Basically find a fruit that has the properties you want, transfer to agar and make a master slant or petri then take some from the master and grow it to agar again for an agar to grain transfer and then to bulk where you pick another fruit to clone and isolate.

But what do you do once your best and final master runs out from continuous bulk grows?  Doesn't cloning a clone result in gene degradation?  How do you keep your preferred isolated fruit alive and healthy?

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InvisibleScrimshaw
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Re: Keeping isolated strains healthy [Re: THK107]
    #23197372 - 05/06/16 11:15 PM (8 years, 8 months ago)

If you put the smallest piece you can to a new dish before you start each grow, you can stretch that slant out for an extremely long time. if you used your master slant to start new grows and are now at the point where you need to create a new slant, that's what I would consider a good problem to have. Best case scenario: put the last piece of original slant to new slant. It shouldn't slow that much if at all. Worst case scenario: make a new slant from spores and maybe find an even better culture. Switching media has been known to prolong cultures, so feel free to experiment with changing your agar recipes too.

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OfflineTHK107
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Re: Keeping isolated strains healthy [Re: Scrimshaw]
    #23197578 - 05/07/16 12:35 AM (8 years, 8 months ago)

Quote:

Scrimshaw said:
If you put the smallest piece you can to a new dish before you start each grow, you can stretch that slant out for an extremely long time. if you used your master slant to start new grows and are now at the point where you need to create a new slant, that's what I would consider a good problem to have. Best case scenario: put the last piece of original slant to new slant. It shouldn't slow that much if at all. Worst case scenario: make a new slant from spores and maybe find an even better culture. Switching media has been known to prolong cultures, so feel free to experiment with changing your agar recipes too.




I found a quote of RR Saying

"Guys, This is old science. It's been done for many years in the commercial mushroom industry. It's the standard way they keep and trade isolated strains of edible mushrooms. They desiccant dry the fruitbodies, then powder them. They store the powder until ready for use, then sprinkle it on agar to regenerate the strain. It stops senescence in its tracks. Try it without soaking in peroxide first. That only weakens the mushroom myc so it doesn't take off as fast. Watch for the first signs of myc growth, and immediately transfer that to a new petri dish. Do this a time or two and you'll have a clean specimen, even from years old dried fruits. It works, trust me. DRIED MUSHROOMS"

Isn't this the same principal as a master slant?

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Offlinecamplo
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Re: Keeping isolated strains healthy [Re: THK107]
    #23197880 - 05/07/16 04:20 AM (8 years, 8 months ago)

:popcorn:


--------------------
:three:

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Invisibleblindingleaf
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Re: Keeping isolated strains healthy [Re: camplo]
    #23197898 - 05/07/16 04:41 AM (8 years, 8 months ago)

you find a clone you like, or an MS isolate you like.  you keep it on a slant, preferably with a wooden stick.  stays in the fridge.  u make copies, like 2-3 if u wanna be careful.  mark the dates on them.  thats ur youngest….keep them safe.  when u want to grow it out, pull out the youngest slant u have instead of cloning from a current grow you have going.

u can look up distilled water storage too.  i don't even know if there is a write up on it, but there are enough posts u can find thru the search to DIY…more so on gourmet board

storage with dry fruits…cool, go for it.  im sticking with slants though…well labeled ones.


--------------------
A few thoughts on cultivation
MICROBIAL HUSBANDRY!!!!

The whole is greater than the sum of its parts

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InvisibleSupalemonhaze
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Re: Keeping isolated strains healthy [Re: THK107]
    #23198032 - 05/07/16 06:54 AM (8 years, 8 months ago)

Quote:

THK107 said:
Basically find a fruit that has the properties you want, transfer to agar and make a master slant or petri then take some from the master and grow it to agar again for an agar to grain transfer and then to bulk where you pick another fruit to clone and isolate.





Let me just correct this for the sake of accuracy. Isolation should not be done on clones and the reason for this is quite simple, anastomosis. When a bunch of strains from a multispore grow are together in a jar or tub, they will eventually fuse together and become one. This means that the good strains will be fused with the bad ones and once this happens, they will not sector away from each other because they have become one.

When you start isolating from a spore inoculated petri dish you are always taking transfers as soon as possible, effectively seperating the strains as fast as possible. This prevents both anastomosis and culture aging. If you see pics and videos of isolation you will see that the growth is never allowed to colonize the whole plate. When you isolate from a clone and end up with a monoculture, there will still be genetics from multiple strains in it. This doesn't happen/doesn't happen often when you start from spores on petris and isolate when the culture is still young.

Also, when you find a good clone and slant it, that will be the culture you always go back to whenever you want to grow it. You do not keep taking clones of clones because the mycelium will start to age, especially if g2gs were made to expand the mycelium.

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OfflineTHK107
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Re: Keeping isolated strains healthy [Re: Supalemonhaze]
    #23198572 - 05/07/16 11:22 AM (8 years, 8 months ago)

Quote:

Supalemonhaze said:
Quote:

THK107 said:
Basically find a fruit that has the properties you want, transfer to agar and make a master slant or petri then take some from the master and grow it to agar again for an agar to grain transfer and then to bulk where you pick another fruit to clone and isolate.





Let me just correct this for the sake of accuracy. Isolation should not be done on clones and the reason for this is quite simple, anastomosis. When a bunch of strains from a multispore grow are together in a jar or tub, they will eventually fuse together and become one. This means that the good strains will be fused with the bad ones and once this happens, they will not sector away from each other because they have become one.

When you start isolating from a spore inoculated petri dish you are always taking transfers as soon as possible, effectively seperating the strains as fast as possible. This prevents both anastomosis and culture aging. If you see pics and videos of isolation you will see that the growth is never allowed to colonize the whole plate. When you isolate from a clone and end up with a monoculture, there will still be genetics from multiple strains in it. This doesn't happen/doesn't happen often when you start from spores on petris and isolate when the culture is still young.

Also, when you find a good clone and slant it, that will be the culture you always go back to whenever you want to grow it. You do not keep taking clones of clones because the mycelium will start to age, especially if g2gs were made to expand the mycelium.




Thanks for the help, so how do you further isate a clone without bringing apon cell degradation?  Would you just need to get lucky early on?

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InvisiblePastywhyteMDiscord
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Re: Keeping isolated strains healthy [Re: blindingleaf]
    #23198596 - 05/07/16 11:29 AM (8 years, 8 months ago)

Quote:

blindingleaf said:
you find a clone you like, or an MS isolate you like.  you keep it on a slant, preferably with a wooden stick.  stays in the fridge.  u make copies, like 2-3 if u wanna be careful.  mark the dates on them.  thats ur youngest….keep them safe.  when u want to grow it out, pull out the youngest slant u have instead of cloning from a current grow you have going.

u can look up distilled water storage too.  i don't even know if there is a write up on it, but there are enough posts u can find thru the search to DIY…more so on gourmet board

storage with dry fruits…cool, go for it.  im sticking with slants though…well labeled ones.




:likeaboss: I'm gonna do what this guy said :derfase:

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InvisibleRaven44
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Re: Keeping isolated strains healthy [Re: Pastywhyte]
    #23198671 - 05/07/16 12:01 PM (8 years, 8 months ago)

I think the answer OP is looking for is the "p value" system. Also "sesenence"

Once u get ur slant made we can call it "p1"

When u bring out ur slant, and make a dish and let it grow out.. this would be "p2". Correct me if I'm wrong here

So u make a transfer from the p2 dish. Now we have a new dish called p3.

Okok, now say ur slants are all used up and need to make new slants... u prob have some dishes hanging out labeled p3 maybe p4.

Sooooo, we take the p4 dish and make a new set of slants... these slants ARE NOT p1 slants. These slants are p4 slants....
(Or would it be p5 pasty lol??)

Anyways, Roger rabbit says secenese isn't an issue until a "p value " of p100 or so

So, look into sesenence op. And also p value

Ur worries should b set to rest after u do

Edited by Raven44 (05/07/16 12:02 PM)

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InvisiblePastywhyteMDiscord
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Re: Keeping isolated strains healthy [Re: Raven44]
    #23198696 - 05/07/16 12:12 PM (8 years, 8 months ago)

P values impact regarding senscence change depending on species. Some can go for more than others. Also p100 assumes that the the majority of the time the culture is refrigerated. You certainly cannot G2G 100 times.

Also P1 would be the first instance of an isolate or master plate. The slant would be P2.

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