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monkey_accident
fail now learn later


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Maintaining favorable genetics?
#26559308 - 03/26/20 02:48 PM (3 years, 9 months ago) |
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So I'm at the stage where I finally moved to agar after a few lucky noob runs with MSS and many many G2G failures. Once I have some genetics on agar that I want: 1)what would be the best method to noc a bunch of grain jars? LC? A2G? G2G with a master jar? 2)after I noc those, how do I keep the genetics going? Is it just a matter of cloning from those fruits or should I keep transferring the original plate?
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rumfor69
Bodhicitta Cultivator



Registered: 08/05/11
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1) I prefer A2G then use that master in G2G
2) it's best if you can preserve the original mycelium network as much as possible. People do slants with craft sticks in them. I vac seal and keep it in the fridge only getting it out to make new dishes. Currently I'm also experimenting with a jar of sterilized Water that has an synthetic filter disk lid setup and just letting wedges from an original dish float in it to keep them moist and alive.
3) Make a bunch of spore prints from your first ever fruits and you can start very close to the top again. Apply all these techniques and you can preserve a strain for a lifetime.
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Strainsfordaze


Registered: 05/10/18
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Once you have genetics you would like to preserve for future inoculations it would be wise to make a bunch of master slants and petri dishes as well as a few LCs that you can fall back on.
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monkey_accident
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Re: Maintaining favorable genetics? [Re: rumfor69]
#26559352 - 03/26/20 03:19 PM (3 years, 9 months ago) |
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Gotcha. I have had trouble doing G2G in the past, not sure if it was technique or bad spawn but I'll have to give it another shot. Are you vac sealing a slant or plate? Can you explain what you mean by start close to the top again? I think you mean the genetics will be relatively similar, maybe only needing a few transfers. Also, last thing, do you take any type of extra precaution when doing G2G, my SAB is somewhat short but long so I just crack both lids and try to shimmy out as much of the grains as I can into the receiving jar in the shortest amount of time.
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rumfor69
Bodhicitta Cultivator



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I vac seal mycelium growing on plates seems how the fridge isn't a clean place really and to keep them from drying out. But they will die after 6 months or so from lack of oxygen so they need taken outta the vac bag and Allowed to grow and bounce back a good amount before being re vac sealed and back into the fridge.
By back to the top again I just mean back to the top of whatever you have. Not that it'd be the same. Imagine a family tree or flow chart that starts at the spores you began with, then branches out with everything you do. Printing and cloning your first ever fruits is the closest to the beginning spores you started with(or "the top")
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c10h12n2o
serial dilutor



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Re: Maintaining favorable genetics? [Re: rumfor69]
#26559498 - 03/26/20 04:16 PM (3 years, 9 months ago) |
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#1 g2g or LC. i usually prefer LC but its riskier if your clean tek and cultures arent on point. fast af with proper LC and grain prep
#2 i love slants, i just revived a bunch of 3+ year old cultures that hadnt been refreshed at all. meanwhile sporeprints from same period are being a pain in the ass to get germination from
i just use the sticks, full strength agar (double the usual, 30g rather than 15g LME), parafilm the lids slightly loose, then throw them in ziplocs and place in fridge
ziploc bags are sterile from the factory (the inside obviously, not outside) so i leverage these in lots of ways. i actually individually bag every plate i do, so i can handle it however i want without getting contams on the outside of the plate
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  C10's Agar Guide + Tips and Tricks | c10's Flow Hood Build Guide "Partial knowledge is more triumphant than complete knowledge; it takes things to be simpler than they are, and so makes its theory more popular and convincing." "Convictions are more dangerous enemies of truth than lies" ― Friedrich Nietzsche
Edited by c10h12n2o (03/26/20 04:17 PM)
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monkey_accident
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Re: Maintaining favorable genetics? [Re: c10h12n2o]
#26559799 - 03/26/20 06:50 PM (3 years, 9 months ago) |
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Gotcha, I actually handle my plates all the time, wrapped in parafilm of course.. and don't get contamination on them.
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c10h12n2o
serial dilutor



Registered: 01/21/15
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Yeah that works, I just overkill everything
Its just One less thing that can go wrong, since theres always the slim chance that contams on the outside of the plate can end up inside when you are working with them
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  C10's Agar Guide + Tips and Tricks | c10's Flow Hood Build Guide "Partial knowledge is more triumphant than complete knowledge; it takes things to be simpler than they are, and so makes its theory more popular and convincing." "Convictions are more dangerous enemies of truth than lies" ― Friedrich Nietzsche
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migraineur
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Re: Maintaining favorable genetics? [Re: c10h12n2o]
#26560816 - 03/27/20 09:33 AM (3 years, 9 months ago) |
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Quote:
c10h12n2o said: #1 g2g or LC. i usually prefer LC but its riskier if your clean tek and cultures arent on point. fast af with proper LC and grain prep
#2 i love slants, i just revived a bunch of 3+ year old cultures that hadnt been refreshed at all. meanwhile sporeprints from same period are being a pain in the ass to get germination from
i just use the sticks, full strength agar (double the usual, 30g rather than 15g LME), parafilm the lids slightly loose, then throw them in ziplocs and place in fridge
ziploc bags are sterile from the factory (the inside obviously, not outside) so i leverage these in lots of ways. i actually individually bag every plate i do, so i can handle it however i want without getting contams on the outside of the plate
How long is a culture good for on agar in the fridge?
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Strainsfordaze


Registered: 05/10/18
Posts: 669
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Re: Maintaining favorable genetics? [Re: migraineur]
#26560823 - 03/27/20 09:38 AM (3 years, 9 months ago) |
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Depends on storage method, thickness of plates, fridge temp, and how often you open the fridge. Variably a few months or so. I’ve revived some agar 6 months. If doing slants it could last years
Edited by Strainsfordaze (03/27/20 09:42 AM)
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migraineur
Geezer


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How can slants last for years where as a plate only lasts for 6 months?
Wouldn't they lose their vigour?
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poisoned
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Re: Maintaining favorable genetics? [Re: migraineur]
#26560955 - 03/27/20 11:10 AM (3 years, 9 months ago) |
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Plates have a huge surface area/volume ratio, hence they tend to dry out. Slants have a big water reserve and won't dry out.
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migraineur
Geezer


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Re: Maintaining favorable genetics? [Re: poisoned]
#26560980 - 03/27/20 11:24 AM (3 years, 9 months ago) |
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I thought the cultures just wouldn't live very long. I remember my LCs losing their vigour so when I was using them I remember them being lacklustre at the 1 year mark or something.
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poisoned
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Re: Maintaining favorable genetics? [Re: migraineur]
#26561142 - 03/27/20 01:02 PM (3 years, 9 months ago) |
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Did you give them some time to "wake up"? From slants, you usually go to agar and only then inoculate. This allows myc to get some food and get ready for that grain.
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c10h12n2o
serial dilutor



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Re: Maintaining favorable genetics? [Re: poisoned]
#26561469 - 03/27/20 04:06 PM (3 years, 9 months ago) |
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Has the 9000 year old honey mushroom colony in oregon "lost its vigor"? Or the decades old cube colonies in the wild?
I think the risk of losing vigor in such short time spans is greatly exaggerated. Sure, it can happen, but I think there are usually other issues at play
Your LC probably contammed, suffocated, starved, or something else.
Slants dont dry out as easy as plates, as was mentioned. It's still a good idea to refresh them every 6 mo or year or so, but I've had great success with slants as old as nearly 5 yrs
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  C10's Agar Guide + Tips and Tricks | c10's Flow Hood Build Guide "Partial knowledge is more triumphant than complete knowledge; it takes things to be simpler than they are, and so makes its theory more popular and convincing." "Convictions are more dangerous enemies of truth than lies" ― Friedrich Nietzsche
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