lichten at HELIX.NIH.GOV
Tue Jul 11 15:43:53 EST 1995
>Hi fellow netters,
>I want to photograph some S. pombe cells, both phase and DAPI-stained. Is
>there a fast and simple method for fixing and permeablizing cells for
>DAPI-staining relative to the procedures used for immunofluorescense? Thanks
>"I'd rather be fission."
>P.S. When trashing the "Hoffman and Winston" protocol for plasmid rescue
>from yeast, please refer to it as the "Smash and Grab" technique. Thank you.
DAPI Staining of yeast nuclei
1. Pellet cells, resuspend in 50 ul water. Add 1 ml ETOH.
Alternatively, fix cells in 50% ETOH ahead of time and chill at -20 C.
2. Add DAPI to 0.1-0.2 ug/ml. Let sit 5 min.
3. Spin out, rinse 2x with water. Resuspend in a small volume
(usually 20-50 ul).
Is that fast enough?
lichten at helix.nih.gov
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