in vitro transcription: low incorporation of label

Hinayana Bawagan bawagan at umdnj.edu
Sat Jun 11 08:53:24 EST 1994


     I am posting this for an acquaintance who mentioned the problem to me. 
He needs to make an RNA probe from a cDNA template by run-off transcription
in vitro.  The RNA made incorporates UTP (then he tried CTP) at a very low
efficiency of 1% while the DNA template of the Maxi Script kit produces RNA 
with a 50% incorporation.  He suspects "rearrangement of some sequences in
the cDNA template during the subcloning which causes early termination of
transcription."  
      What do you think?  Thank you.



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