Cleaning up Transcriptions

[brett]

>Does anyone know if ethanol precipitation removes unincorporated
>ribonucleotides from transcription reactions?  If not, how can this be
>achieved?
>
>MEM

Depends on the salt you add. NH3oAc is better at removing unincorporated rNTPs,
but it will still take 2-3 sequential precipitations. Another option is
to use a spin-column or other resin-based purification method. For RNP probes
I usually just cut my full-length transcript out of a sequencing gel, which
gets rid of unincorporated nucleotides as well as abortive transcripts (which
can be suprisingly abundant).


Brett Lindenbach
    
Program in Immunology                              
Washington University - St Louis                  
brett at borcim.wustl.edu