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dodgem
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Question about Generations of 'Strains'
#19461389 - 01/23/14 08:23 AM (10 years, 11 months ago) |
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So I have an inquiry about generations of 'strains'. Lets say you start with a spore syringe and get some growth on agar. From the agar you inoculate a WBS qt jar. All goes well and colonizes 100%, and becomes your G1, correct?. You take that and G2G with other WBS qt jars, which are in turn G2. Say you do this and go up to G4. Now say you take a grain from G4 and put it in a petri dish with some agar and such. Is the growth on agar still considered G4, or is it now G5, or does it some how reset and go back to a G1. Or is the only way to start back at G1 by using spores start over? Or what about a tissue sample from a mature mushroom?
The reason I ask is in the past I would PC 7 jars at a time. I would use a master jar to G2G all 7. From those 7 I would use 5 for a tub, and then the other 2 to G2G with my next two batches of 7 jars, and so on. So pretty quickly I was up to G4/G5, if not further. My tubs are now getting one good flush (5-8 oz dry) and then contaminating a couple days after I harvest. I have not had a tub make it to the second flush in a few months. My guess is that my 'strain' became less contaminate resistant due to all the transfers. So I am hoping to figure out how to start back over with a master jar at G1.
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PirateSwazey



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Re: Question about Generations of 'Strains' [Re: dodgem]
#19461441 - 01/23/14 08:34 AM (10 years, 11 months ago) |
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It would be G5.
That is the point of making a slant so you always have a first gen culture to go back to 
Tissue samples will create the next generation, not a new sequence (it's the same mycelium)
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dodgem
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Re: Question about Generations of 'Strains' [Re: PirateSwazey]
#19463320 - 01/23/14 04:07 PM (10 years, 11 months ago) |
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Does this 'don't go past G3' apply only to G2G's? So say if you were to put some spores on agar, and then over a couple weeks make a few transfers onto new agar plates 3 or 4 times. Does that lead to a G4? Would that also cause it to become more susceptible to contamination?
I guess I will plan on taking a few spore prints from my next couple tubs and try to restart with a new master. I will have to look into this master slant business, have only seen it mentioned a couple times on the boards.
I bought a treasure coast syringe over two years ago, and have just gone from syringe --> LC --> WBS --> agar --> then random WBS and agar for over a year. So I guess that may be a big reason why I kept getting bacterial contams 6 months ago...
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cronicr



Registered: 08/07/11
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Re: Question about Generations of 'Strains' [Re: dodgem]
#19463327 - 01/23/14 04:09 PM (10 years, 11 months ago) |
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thats not how generations are defined, in order to get to your next generation you must fruit the original, all g2g does is expand the original gen much like beer makes you fat
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PirateSwazey



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Re: Question about Generations of 'Strains' [Re: cronicr]
#19463361 - 01/23/14 04:18 PM (10 years, 11 months ago) |
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Quote:
cronicr said: thats not how generations are defined, in order to get to your next generation you must fruit the original, all g2g does is expand the original gen much like beer makes you fat
I have heard some people say that a new generation doesn't start until you take a sample from a fruited clone but I got my info from Stamet's book 
Quote:
dodgem said: Does this 'don't go past G3' apply only to G2G's? So say if you were to put some spores on agar, and then over a couple weeks make a few transfers onto new agar plates 3 or 4 times. Does that lead to a G4?
No agar transfers don't make up a generation 
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cronicr



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Re: Question about Generations of 'Strains' [Re: PirateSwazey]
#19463371 - 01/23/14 04:20 PM (10 years, 11 months ago) |
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yeah he defined it wrong but it's not the biggest of issues just an odd way of keeping track
-------------------- 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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PirateSwazey



Registered: 12/12/12
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Re: Question about Generations of 'Strains' [Re: cronicr]
#19463385 - 01/23/14 04:24 PM (10 years, 11 months ago) |
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I dunno cronic,
I have experienced cultures losing their vigor after multiple generations of G2G transfers...
That's why I got into working with agar in the first place...
so I would always be able to have a clean, fresh generation to start with
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vikingsc
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Re: Question about Generations of 'Strains' [Re: PirateSwazey]
#19463404 - 01/23/14 04:30 PM (10 years, 11 months ago) |
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Quote:
PirateSwazey said: I dunno cronic,
I have experienced cultures losing their vigor after multiple generations of G2G transfers...
That's why I got into working with agar in the first place...
so I would always be able to have a clean, fresh generation to start with 
That's not what he's saying. Generations just isn't the correct term for it is what he's saying. A generation is technically a spore print fruited and back to a spore print.
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cronicr



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Re: Question about Generations of 'Strains' [Re: vikingsc]
#19463427 - 01/23/14 04:34 PM (10 years, 11 months ago) |
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This is incorrect. Don't confuse grain to grain transfers, etc., as generations even though a well-known book got it wrong 30 years ago.
The op had it right the first time. Spores to fruits constitute a generation. You are the next generation from your parents. Your kids will be the next generation from you.
If you're skinny and then get fat(expanded) it isn't a generation, it's beer. RR http://www.shroomery.org/forums/showflat.php/Number/17002119#17002119
-------------------- 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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vikingsc
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Re: Question about Generations of 'Strains' [Re: cronicr]
#19463449 - 01/23/14 04:39 PM (10 years, 11 months ago) |
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I was reading a different thread the other day of RR correcting someone and saying what the actual correct term for the next g2g was, but I can't remember or find the thread.
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PirateSwazey



Registered: 12/12/12
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Re: Question about Generations of 'Strains' [Re: vikingsc]
#19463520 - 01/23/14 04:56 PM (10 years, 11 months ago) |
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vikingsc
Mushroom Enthusiast



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Re: Question about Generations of 'Strains' [Re: PirateSwazey]
#19463535 - 01/23/14 04:58 PM (10 years, 11 months ago) |
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Hahaha
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dodgem
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Re: Question about Generations of 'Strains' [Re: vikingsc]
#19466515 - 01/24/14 08:18 AM (10 years, 11 months ago) |
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So now that I have all that cleared up, does taking a grain and putting it on agar 'clean/refresh' it or anything. Is there anyway if working with a long time culture to get back its vigor? Or should I just spore print and start over, making sure to keep a master slant to begin with?
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bodhisatta 
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Re: Question about Generations of 'Strains' [Re: dodgem]
#19466571 - 01/24/14 08:43 AM (10 years, 11 months ago) |
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Quote:
dodgem said: So now that I have all that cleared up, does taking a grain and putting it on agar 'clean/refresh' it or anything. Is there anyway if working with a long time culture to get back its vigor? Or should I just spore print and start over, making sure to keep a master slant to begin with?
taking a grain back to agar is like doing a g2g to agar the mycelium is still expanding and getting older, not a new generation just older and fatter. Your best slants will be ones made from spores to agar and then how ever many transfers to get a culture you want be it an isolate or still multiple genetics, fruit it out and save the cultures that performed well, now your slants have very "fresh" genetics on them which will always be very close to the origin spores rather than old and fat. But then again you can get away with slanting older genetics as many many many people do but it's not something you would likely see a commercial farm doing.
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