DNA/b-gal staining

Michel Leroux mleroux at unixg.ubc.ca
Sat Mar 25 15:29:33 EST 1995


In article <9503231826.AA06819 at brenner.Scripps.EDU>, rakow at SCRIPPS.EDU
(Terese Rakow) wrote:

>         I am looking for a simple method of staining worms with Hoechst
> after b-gal or antibody staining.  Can they just be mounted in PBS-Hoechst
> or does it have to be washed out?  Also has anyone done b-gal, antibody and
> DNA staining in the same specimen?  Any advice would be welcome.
>         Thanks,
>         Terese Rakow <Maruyama lab>
>         The Scripps Research Institute

You can include DAPI stain, for example, in your mounting solution when
you do antibody staining. This allows you to visualize your antibody
stain, switch filters, and then visualize DNA (DAPI) stain. So, you can do
both at once. For b-gal, I think that you should be able to take your
X-GAL stained worms, put them in mounting solution containing Hoechst or
DAPI DNA stains, and visualize using the appropriate filter. I don't a
washing step would be necessary since Hoechst/DAPI stains nuclei very
intensely with little background.

Michel Leroux
mleroux at unixg.ubc.ca



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