RNA Probe woes

Carlisle Landel landel at helios
Mon Sep 19 13:34:55 EST 1994


In article <35jnec$lk9 at cc.tut.fi>, Jukka Luoma <bljulu at uta.fi> wrote:
>D.Greeve <DAEMON> (darreng at ccmar.csiro.au) wrote:
>: Hi netters.
>
>:    I am currently experiencing difficulties in producing P32 labeled RNA 
>: probes from a "Bluescript KS+"  plasmid. I am currently using a kit marketed 
>: by "Bresatec" (- an Australian company), designed to do this, but I am getting 
>: almost no transcription using T7 and my plasmid. The kit control appears to 
>: work, however with identical conditions, my plasmid does not.
>
>: Any ideas on the matter would be much appreciated....
>
>: Darren Greeve.
>
>I have had similar problems and usually the cause of the promlems was too
>low plasmid concentration. Are you sure about your plasmid concentration?
>Your plasmid concentration should be about 0.1ug/ul in the final volume.
>
>Jukka


Another common source of problems is RNase in your plasmid prep.  

Carlisle




More information about the Methods mailing list