Standardising Northerns

Ola Myklebost olam at radium.uio.no
Thu Apr 15 11:18:46 EST 1993


In article <1993Apr13.152808.8716 at gserv1.dl.ac.uk>, sbhattac at crc.ac.uk (Dr.
S. Bhattacharya) wrote:
> 
> Hi all,
> 
> I read somewhere that a 18S rRNA probe is a better option than the usual
> actin for correcting loading variations on northern blots.  Does anyone have 
> any ideas on this,
> and has anyone done it using a oligonucleotide probe to 18s RNA (if so could 
> they let me know the oligo sequence), as I do not have a cDNA probe.  

We use 

5' GCG CGA GAT TTA CAC CCT CTC C for at least human and rodent 28S rRNA.

Hybridisation is done in "Church hyb" at 55 C and washed in 40 mM Na
(phosphate) 1% SDS at 50 C.

The good thing with oligos is that they are easy to get off again. A
problem might be the excess of target on the filter if you hybridise in
stationary bags. 


Ola Myklebost             Email olam at radium.uio.no
Dept of Tumor Biology
Inst for Cancer Research
The Norwegian Radium Hospital
N-0310 OSLO, Norway
Tel +47-22-50-60-50, xtn 9830, Fax +47-22-52-24-21



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