Help: In vitro Transcription

Lars Thelander lars.thelander at medkem.umu.se
Tue Jun 13 07:05:27 EST 1995


I am trying to start up an in vitro transcription system to study a particular mammlian 
promoter. I have a couple of questions:

(1) Is there a good protocol for measuring the amount and length of in vitro synthesized RNA 
(using nuclear extracts) using an 35S-labeled riboprobe? Or is it better to use a 32-labeled 
probe?

(2) Any suggestions on protocols for run off transcription (from a linearized plasmid) with 
labeling the RNA (again using nuclear extracts)would be most appreciated.

Thank you.


Sincerely,

Eric Hanson




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