🚀 go-pugleaf

Hi Don,

We understand your sadness about absence of UV laser... How nice and easy
would be our life if we could counterstain nuclei with DAPI...

You did not write in which cells - live or fixed - you need to stain DNA. I
can help you with fixed material:

We routinely use two ways of DNA staining for confocal. Both of them,
however, require RNase treatment (could be done beforehand or during
incubation with antibodies). RNasing is done at 37C, 200mg/ml in 2xSSC (or
PBS). Then we use:

(1) PI, propidium iodide, 1 - 10 mg/ml, in 2xSSC or PBS; it is important to
remember that PI stains denatured (ss) DNA  better than dsDNA, therefore
undenatured nuclei should be stained MUCH longer (say, 1 min for denatured
DNA, and 15 min for native DNA in human cells).
or
(2)YOYO (Molecular Probes) at 2.5 nM in 2xSSC or PBS. We prepare fresh
working solution from a stock, 2.5mM in DMSO. Human nuclei in 3D cells are
stained  20 - 30 min.

Irina.

===========================
          Dr.I.Solovei

   Institute of Anthropology
       and Humangetics
   University of Munich (LMU)

    Richard-Wagner-Str. 10/I
    80333   Munich   Germany
===========================
irina.solovei@mail.lrz-muenchen.de
      tel.+49 (0)89 2180 6713
      fax+49 (0)89 2180 6719
===========================