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Marguerite Evans p2158740 at ACSUSUN.ACSU.UNSW.EDU.AU
Fri Jun 14 23:06:53 EST 1996


Hi Isidro,

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=
=20
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!!

Marguerite Evans
School of Biochemistry and Molecular Genetics
UNSW  Sydney  2052
NSW
Australia

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.
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>     I need to find protocols for Northern and ribosomal RNA electrophores=
is=20
> courrently working with yeast and I have tecnicals troubles.
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>    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.
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>    I heve also stain the agarose gels with etidium bromide and I have not=
=20
> seen the bands 25S and 18S rRNA.
>    All my glassware has been trated with DEPC.
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>=20
>    I shall be gratefull for any help.
> =20
>=20
> * 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
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>=20




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