BRL RNA ladder hybridizes to my probe
ojar.melefors at postmac.cmb.ki.se
Fri Aug 18 09:16:43 EST 1995
In article <DDH3B2.FEr at ns1.nodak.edu>, stallard at badlands.NoDak.edu (Lana C
>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
The same thing happens to me with many of the probes I use.
I presume that just a high enough concentration of the RNA weight standard
will cause cross-hybridisation with almost any probe if the filter is not
washed stringently (same as for rRNA).
But cross hybridisation makes me happy :-) as it facilitates the correct
determination of band sizes!
"Speaking for myself"
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