help: in vitro transcription

brett brett at BORCIM.WUSTL.EDU
Tue Jun 13 13:43:04 EST 1995


>Does anyone know of a good reference for RNase protection assay to measure the
>amount of RNA in 
>from an in vitro transcription assay? Can this readily be done by making a
>riboprobe (off a T7 
>promoter) and labeling with 35SUTP (or another rNTP) or do I need to use 32P?
>Also, I am 
>planning to label RNA with 32P off a linearized plasmid with my promoter of
>interest but am 
>open to alterantives. I am just starting in vitro transcription so any help
>would be most 
>appreciated.
>
>Thank you.
>
>Eric Hanson


It depends on what you wish to use your RNA for. In our lab, we simply spike
the transcription reaction with 3H-UTP, then measure the amount of incorporated
3H, and calculate the yield of RNA, with good quantitative results.



Brett Lindenbach
    
Lucille P. Markey Student in Human Pathobiology    
Program in Immunology                              
Washington University - St Louis                  
brett at borcim.wustl.edu                             






More information about the Methods mailing list