BRL RNA ladder hybridizes to my probe
poets at ccmail.orst.edu
Thu Aug 17 17:56:43 EST 1995
You've got vector sequence contamination. Try generating your probe
with internal PCR primers. BRL's got a PCR labelling kit which works
well. Use a very, very, very small amount of template in the reaction.
stallard at badlands.NoDak.edu (Lana C Stallard) wrote:
> I wonder if anyone has had this happen? I am using Gibco BRL RNA weight
> standards (made from phage lambda DNA spliced to an RNA transcript). I get
> significant hybridization with a mouse DNA probe (a cDNA probe of
> angiotensin converting enzyme) to the ladder when i transfer the ladder
> onto a Northern blot. Why would there be so much sequence similarity between lambda and a mouse gene? or could my probe still contain some sequences of the vector it was produces in (Bluescript
> plasmid) which is similar to the phage DNA? My boss is convinced i have a
> very non-specific probe but i think there must be a reason.
> Thanks for any comments or ideas you may have.
> Lana Stallard
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