I've pelleted down the overnight ampicillin supplemented cultures of DH5ALPHA for pET22b isolation 5 days ago and stored them on -20°C without adding any buffer or glycerol. Would the cells be viable for mannual plasmid isolation?
Dear Syeda Batool
viable NOT, do not try to re-hrowth. IT will fail.
Howeve if the amount of frozen cell pelletted is enough you can try to resuspend direclty with lysii buffer suitable for mini kit and perform plasmid extraction. In case you are able to exctrasct something you can trasform it again in fresh cells, propagate and store the new cell batch.
best
Manuele
I've frequently done mini preps from frozen cell pellets, it should be fine. However you might have a bit more genomic contamination than usual because some cells may lyse with the freeze thaw cycle. But probably not enough to be a serious problem.
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