smear Northern

[J. David Spafford]

In Article <199608070043.JAA24219 at sesame.nibb.ac.jp>, kankaku at NIBB.AC.JP
(kankaku) wrote:
>I believe I am not poor at Northern.  However, this time, I have trouble with 
>obtaining signals for my rare transcript.  I tried various things, using
>DNA probes, 
>riboprobes, partial digestion, total RNA, polyA RNA, formaldhyde, glyoxal, 
>various membranes, various hybridization solutions.  But, nothing improved 
>the smear, but specific band.  Of course, my RNA sample looks very intact 
>because signals for other transcripts as well as rRNA are beautiful. I quess 
>the turn-over of the rare mRNA is very rapid.  I need to know the size of the 
>mRNA.  So, other methods such as RPA are not useful for my purpose.  
>I would appreciate it if someone would suggest how to obtain a good 
>Northern signal for such a transcript.  

If your RNA is not degraded, then it may be your probe.  Try a S. blot.  If
you get a smeary result, you have your answer.


J. David Spafford
Department of Biological Sciences, University of Alberta,
Edmonton, Alberta.  T6G 2E9   EMAIL: jspaffor at gpu.srv.ualberta.ca