Short oligo probes for Northerns ??
Claudia Sutton
cas9 at cornell.edu
Tue Apr 16 05:44:09 EST 1996
In article <31735013.458E at rmit.edu.au>, Patrick HJ Falckh
<p.falckh at rmit.edu.au> wrote:
> 'lo molbolians,
>
> I was wondering if anyone has had experience in using short oligos
> (20-25 base) to probe Northerns. I've only previously used 250-500
> base probes and was wondering if the shorter probes would function
> adequately.
>
> I'm considering using end-labelled PCR primers (20-25 bases) to do the
> probing , which are degenerates for a class of enzymes; but would it
> work.
>
> Your input, and experience, would be most appreciated.
>
> Thanks in advance,
>
> --
>
> Patrick HJ Falckh PhD # #
> Key Centre for Applied & Nutritional Toxicology # # # # #
> RMIT - City Campus # Q Q #
> GPO Box 2467V, Melbourne Vic 3001 # o #
> # ~ #
> Telephone : 03 9660 3136 ##<->##
> Fax No. : 03 9663 6087 # #
> Email : p.falckh at rmit.edu.au # # # #
> ## ##
> "No matter how tall your grandfather # \/ #
> was, you have to do your own growing" # /\ #
> ooO Ooo
Patrick,
I've used 25 mers, but they were not degenerate. Also, my primers were
antisense to mitochondrial transcripts, and the RNA was from purified
mitos. What the heck, give it a try, but be sure to use nitrocellulose as
the membrane.
My paper was in MCB, Sutton et al, in 1994 I think.
Claudia
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