problems with background doing in vitro transcription/translation

jlight at SICKKIDS.ON.CA jlight at SICKKIDS.ON.CA
Fri Jun 28 12:35:09 EST 1996


I'm using the TNT coupled reticulocyte lysate system from Promega for in
vitro transcription/translation, and I'm getting a lot of background signal
at the top and bottom of my autorads.  I can understand that the signal at
the bottom would be unincorporated 35S-methionine which I can run off the
gel.  However, why the signal at the top?  The reason that this is a
problem is that my product has a high MW and is obscured by the background
at the top.  Also, I encounter background smearing in many of my sample
lanes, although the positive control lane looks nice.  Any advice on how to
get rid of background with this assay would be appreciated.  Thanks in
advance. Jeff.





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