Hello Kari,

If we want to stain nuclei only we do not fix the cells at all. We just wash
the cells once with buffer, add 1 µg/ml DAPI, incubate 5 min and directly
observe fluorescence.

Cheers,

Sascha.


Am 10.11.2011 16:40 Uhr schrieb "Kari Naylor" unter <[log in to unmask]>:

> Dear Dicty colleagues
> 
> I am writing to see if anyone can help us on two disparate issues.
> 
> 1) DAPI staining.  We'd love some success stories on DAPI staining.  We've
> fixed the cells as recommended (in 3% glutaldehyde- maybe this is the problem)
> but the staining is quite poor. Any suggestions would be appreciated.
> 
> 
> 2) we are trying to make a knockout strain and are having some significant
> issues attaching our up and down stream genomic pieces to BSR gene.  We have
> decided to try pricing out some companies that synthesis large pieces of DNA,
> but to do that we need the entire sequence of the BSR gene (we're using
> pUCBsrdeltaBam).  Does anyone have the complete sequence???
> 
> 
> Thanks in advance
> 
> Kari
> 
> Kari Naylor PhD
> University of Central Arkansas
> Biology Department
> 139 Lewis Science Center
> ph:501-450-5826
> [log in to unmask]

-- 
Dr. Sascha Thewes
Institut für Biologie - Mikrobiologie
Fachbereich Biologie, Chemie, Pharmazie
Freie Universität Berlin
Königin-Luise-Str. 12-16
D-14195 Berlin

Tel.: 030-838-53373
Fax: 030-838-57773
Email: [log in to unmask]
Web: http://www.biocircle.fu-berlin.de/mikrobio2/index.php