Monitoring RNA synthesis

Neil Winegarden nwinegar at erin.utoronto.ca
Mon Feb 26 09:56:01 EST 1996


My apologies if my first post did make it out, however since I received 
no replies I thought I'd try again.

I want to measure the synthesis of RNA inside of cells.  Specifically, I 
would like to be able to quantify and localise the amount of transcript 
prepared inside of cells, and I need to perform the localisation by 
immunofloresence.  Normally I would used tritiated uridine to measure the 
amount of synthesis but the resolution of localisation is not high enough 
for my needs.  I was thinking of using something like DIG-UTP however my 
understanding is that this molecule is too large to get into cells.  If 
at last resort I must I could microinject this molecule, however this 
would make my task much more tedious than I hope it to be.

Specifically my question is:  Does anyone know of some labelled precursor 
that would be taken up by cells?  Would DIG-uridine be taken up?

Any help would be greatly appreciated.

Thanks in advance,

-- 
Neil Winegarden
University of Toronto - Zoology
nwinegar at credit.erin.utoronto.ca



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