Protocol for DAPI staining yeast

ZX xuez at esvax.dnet.dupont.com
Mon Feb 17 09:27:23 EST 1997


James R. Bone wrote:
> 
> Is there and existing protocol for simply DAPI staining of yeast for
> visualization under fluorescence optics (no immuno, just DAPI).
> 
> Thanks,
> 
> Jim

Fix with ethanol and stain with DAPI.  It is that simple.  To fix, add 
two volumes of ethanol to one volume of cell culture.  Let stand at 
room temperature for 30 to 60 minutes.  spin down and wash a couple of 
times with your favorable buffer.  I usually stain with a 1:2000 
dilution of 1 mg/ml DAPI stock solution.  If your cell is fresh and 
under O.D. 0.8 to 1.0, you can also stain without fixing.  You will 
notice that some cells won't stain well.  cells off a fresh plate can 
be resuspended in YFB and stained too.  It is actually better than the 
liq. culture if you stain without fixing.  Lastly, you can also fix as 
in Immunoflurescence protocol and stain.  You need to spheroplast for 
good result if you do this.

Good luck.

-- 
Zhixiong Xue
DuPont Central Research
E402/4237
P.O.Box 80402
Wilmington, DE19880-0402
(302) 695-9465



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