Bad message with Ambion Message Machine

[Alan Neely]

While most of the time I obtain high yield of a single length message
transcribed with Ambion Kit and then express functional protein in 
Xenopus oocytes some time I obtain a smear. This happen with certain 
clones and it is not due to RNAse contamination. We tried changing the
ratio cap/gtp, concentration of template and temperature. Absorvance 
reading show a ratio of 1.75 to 1.87 and yield in the 19 to 20 
micrograms... (The same clone transcribed with Promega give a clean
ban of the right size but the yield is to low and expression in oocytes
unaccetable). 

Any similar experiences and posible solutions ??


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Alan Neely, Ph.D.  
Texas Tech U. Health Sciences Center
Lubbock TX, 79430