secondary bands after in vitro transcription

schmitz jschmitz at mail.mpiz-koeln.mpg.de
Wed Apr 15 05:29:22 EST 1998


Hello!
I have a problem with my latest in vitro transcription assays.
After in vitro transcription of some DNA templates using T3 or T7
polymerase
I detect additional bands on a denaturing agarose gel. These bands are
smaller
than the expected product and thus should be due to prematurely
terminated transcripts. 

Is there any protocol available to avoid premature termination during in
vitro transcription? 

Is it possible to add compounds like DMSO into the in vitro
transcription assay
in order to melt RNA secondary structure and thus avoid premature
termination? 

I will greatly appreciate any suggestion to overcome this problem.


Thanks JuerGen

(JuerGen is best expressed at 10 p.m in 10x beer)



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