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bongoman
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Agar strategy: expanding from germ plate > T1 > T2 etc
#26411041 - 01/01/20 02:25 PM (4 years, 1 month ago) |
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Let’s say I have 4 pretty clean looking germ plates.
I’m ready to make my first transfer - if all germ plates look equal in growth, cleanliness etc, would you transfer from all 4 plates?
Let’s call the next generation of plates T1 - how many T1 plates do you typically aim to create from your germ plates?
Is it enough to just make a couple of transfers from each plate? Or make more?
I’m not looking to isolate, just to get good clean orderly MS growth so I can go to grain at around T4.
Likewise when going from T1 to T2? How many plates should I make?
I’m a little confused on how many plates to create from a donor plate on the way to getting to the point where I can grow to grain?
Edited by bongoman (01/01/20 02:34 PM)
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A.k.a
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Re: Agar strategy: expanding from germ plate > T1 > T2 etc [Re: bongoman]
#26411047 - 01/01/20 02:31 PM (4 years, 1 month ago) |
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Less than you think.
I just started recently with agar and have a stupid number of plates from the clone I first used.
I would just take one or at most two T1s. Maybe it’s different from spores than clone but most of my T2s seem good for grain, and def by t3.
My biggest mistake was taking too many T1s my first time, you really only need one and you can take more than enough transfers from it. Now I just have a bunch of unused T1 plates sitting around hoping they pin.
The only time I had plates that were totally unusable was pin mold but I only got that trying to clone stumps days after a harvest. As long as your technique is decent you really shouldn’t lose much to contams.
My strategy at this point is clone>one T1 then if it looks even slightly sketchy I’ll take one t2. From that point I’ll take as many transfers as I’m going to have jars.
For example I’m going to do a 4qt grow of Malabar next and had a clean t2 plate. So I took four transfers off that plate and each will grow out and go into a jar.
Im still learning though so this is just where I’m at now that I’m not brand new, and being confident in my SAB skills is a big part of it. Before I would always want back ups Incase something went wrong.
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LAGM2020     
Edited by A.k.a (01/01/20 02:43 PM)
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bongoman
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Re: Agar strategy: expanding from germ plate > T1 > T2 etc [Re: A.k.a]
#26411051 - 01/01/20 02:36 PM (4 years, 1 month ago) |
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Thanks, that makes sense. I figure I can always expand to more plates as I’m ready to go to grain.
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vc77
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Re: Agar strategy: expanding from germ plate > T1 > T2 etc [Re: bongoman]
#26411058 - 01/01/20 02:41 PM (4 years, 1 month ago) |
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Take as many transfers as you want/need. Things exponentiate pretty quick.
If you’re sure your T1s are clean you can take them to grains. I almost always run T2s from MS. Sometimes T3s, almost never T1s.
I guess it also depends on how you choose to inoculate. If you’re equating each transfer to a step in cleanup/isolation you can further isolate by way of inoculation. If you take a choice wedge from your T1 plate to LC or grains that’s essentially a T2 colonizing your grains. If you use the entire plate then it isn’t.
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