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MycoWeek
Elementary Student




Registered: 07/05/21
Posts: 674
Loc: USA
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How many "validation" grow from MS do you run before you settle on a keeper?
#27570520 - 12/05/21 03:44 PM (2 years, 5 months ago) |
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I am abit hung up on how to proceed with isolating a good phenotype from the varieties I am working with, greatly appreciated if you can help me answer some questions below.
1. If you germinated a plate of multispores from syringe or print, how many agar plate do you transfer to for first transfer? out of all those plates, how do you pick the section for 3rd transfer and so on?
2. By the time you hit 3rd transfer, what are the average number of plates you have for that variety?
3. how many "validation" grows do you run once you have good mycelium on agar?
4. at what point of the process do you go "backwards" to start off from fresh spores, or earlier transfers?
5. If you have gone backwards, were you able to find better phenotype?
6. can performance on agar be deceiving or inconsistent, meaning it's not the fastest performer on agar, but performs better once transferred to grains or spawned to substrate?
Thank you all in advance.
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SingularFusion


Registered: 10/31/18
Posts: 1,228
Last seen: 1 month, 22 days
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Re: How many "validation" grow from MS do you run before you settle on a keeper? [Re: MycoWeek] 3
#27570902 - 12/05/21 10:40 PM (2 years, 5 months ago) |
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from a single MS plate, I'd maybe make another six or something personally, then throw half of them once they are done, choose the best three. Make another six from those three for the T2, throw three worst again, use the best plate of the remaining three to noc some grain and to make a spare plate at the same time so you can rinse and repeat if desired or needed. Keep the other two good plates around and work on them further and keep making grain from them and their transfers, making more tubs in the coming weeks.
I personally space them out so I am not overwhelmed with tubs all at once, I only run three or four in constant rotation. Still this allows a lot of work to be done.
Probably I would say mostly I can grow several tubs worth of prospecting material with a 20 sleeve of plates using the transfer schedule example above. Sometimes it takes a little more if there is a T3 needed before going to grain, but often the T2 series is good looking to me.
From that prospecting material, which may be like 4 - 8 tubs worth, I will choose a clone. Clone it. Do some small agar on it, just to ensure cleanliness. Then run it hard to see if it is really what I hoped to capture when I cloned it (sad part for me, it often isn't). If it is what I hoped it would be, then I make some prints, and reclone it, even though people say not to do that. Once the reclone looks good on agar I make a slant.
So far the things I have kept I have not gone back to the print from the clone and attempted to stabilize the genetic line, I've only been at this a couple of years and am a little obsessed by finding clones and slanting them at this point it seems
I find personally that agar can be deceiving... things can look weak and whispy and perform spectacularly once on grain. I have a few nice clones that make quite whispy light myc and fruit very well. The key is to not obsess yourself with rhizo IMHO. It looks good on a plate but I find zero correlation between rhizo and fruiting performance personally
hope that managed to help you man
all the best 
Edit: And of course I didn't mention that the whole cycle may be interrupted at any point where I decide it is all junk and I throw everything in the bin and start again from the original spores to hope for a better roll of the dice
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Edited by SingularFusion (12/05/21 10:43 PM)
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MycoWeek
Elementary Student




Registered: 07/05/21
Posts: 674
Loc: USA
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Re: How many "validation" grow from MS do you run before you settle on a keeper? [Re: SingularFusion]
#27571006 - 12/06/21 03:51 AM (2 years, 5 months ago) |
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Quote:
Nef said:
Great post! this is exactly what I am looking for, I am going to start following your protocol so I can keep numbers of petri dishes under control.
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xVadisx
MountainsDogsAndFungi



Registered: 09/03/21
Posts: 1,324
Loc: Secret World HQ
Last seen: 4 hours, 32 minutes
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Re: How many "validation" grow from MS do you run before you settle on a keeper? [Re: MycoWeek]
#27582067 - 12/15/21 07:36 AM (2 years, 5 months ago) |
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Things are getting quickly out of control for me here -- I'm realizing the amount of oats/bags/tubs I'm about to have running if I don't want waste. I guess not a bad problem to have -- but I think I need to start being more selective moving forward.
I'm very curious about the going back to spore part...
-------------------- Thanks to everyone for sharing what they know . 165F for 24hrs. Better Holes Tek (Use a Step Drill Bit) Don't forget your towel: THE HITCHHIKER'S GUIDE TO THE SHROOMERY
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