I personally am still having difficulties with Northerns, but I think=20
mine is primarily a probe-related problem, but others in the lab have had=
success with the protocol in "Methods in Yeast Genetics" A Cold Spring=20
Harbour Laboratory Course Manual. 1994 edition by C. Kaiser, S.=20
Michaelis and A. Mitchell.
Be sure your calf thymus DNA (or herring sperm DNA) is sheared well. It=20
needs to be small. Good luck!!
School of Biochemistry and Molecular Genetics
UNSW Sydney 2052
voice: (+61)(2)385 2030
fax: (+61)(2)313 6271
email: p2158740 at acsusun.acsu.unsw.edu.au
On 14 Jun 1996, Isidro Contreras Morales wrote:
> Hi everyone.
> I need to find protocols for Northern and ribosomal RNA electrophores=
> courrently working with yeast and I have tecnicals troubles.
> I have run electrophoresis 5, 10, 15 ug of rRNA was extracted with:
> a) acidified phenol, heating to 65=A7 C.
> b) phenol-clorophormo- isoamilic and glass beads(bortex).
> and cuantified by OD at 260 nm.
> I heve also stain the agarose gels with etidium bromide and I have not=
> seen the bands 25S and 18S rRNA.
> All my glassware has been trated with DEPC.
> I shall be gratefull for any help.
> * Isidro Contreras Morales * Email: a73055 at uachih.uachnet.mx> * Division de Estudioos de Posgrado =20
> * Facultad de Ciencias Quimicas * Tel. (14) 13-18-28 Ext.117 =20
> * Universidad Autonoma de Chihuahua * Fax. (14) 14-44-92
> * Apartado Postal 1542-C