You mean a one month old plate with colonies that you left on the fridge? It might work, but most likely won't.
The only clean solution is to re-transform and prepare a working cell bank (i.e. glycerol stocks), then use a fresh aliquot every time you need to perform an expression experiment. Some people with plenty of lab hours even claim that with T7-based plasmids you need to retransform every time, although that is not my personal experience (in my hands, it has only happened in a repeatable and consistent manner with very specific constructs).
Hope it helps!
You mean a one month old plate with colonies that you left on the fridge? It might work, but most likely won't.
The only clean solution is to re-transform and prepare a working cell bank (i.e. glycerol stocks), then use a fresh aliquot every time you need to perform an expression experiment. Some people with plenty of lab hours even claim that with T7-based plasmids you need to retransform every time, although that is not my personal experience (in my hands, it has only happened in a repeatable and consistent manner with very specific constructs).
Hope it helps!
yeah i mean old plate with colonies and i have T7 based plasimd
These strains usually have some recombinase activity then you can experiment problems with the protein overexpression and purification. If the transformation process is not time consuming I would reccomend you to do it again but of course the most plausible situation is that the old plate work perfectly fine.
how are you
if have no choice but to use that plate
subculture them into LB broth then make another master plate out of new cultured colonies, it will make bacteria a bit healthier and might want to use less than normal antibiotics
make sure both plates and LB broth are at room temperature ( prioritize LB broth )
but the best strategy is that you do every step as fast as u could in like a week, the fresh the better
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