Total RNA quantitation

Colin Rasmussen crasmussen at anat.med.ualberta.ca
Thu Jun 8 14:29:46 EST 1995


In article <3r7a24$sgp at yama.mcc.ac.uk>, @fs1.ho.man.ac.uk (hfakhour) wrote:

> Hello
> 
> I am extracting total RNA from a small number of cells (< million)  using 
> Ultraspec RNA reagent. The yield is too low to be measured accurately by 
> spectrophotometry.Does anyone have any suggestions for accurate 
> measurment of small amount of total RNA concentration (< 5ug)
> to be used for quantitative RT-PCR? Help is very much  appreciated.

If you are most interested in mRNA, you could probe a fraction of your
sample with and end-labelled oligo-dT.  The best way would be to use a
slot-blotter, and then cut out the bands and count directly, or if you
have access to a phosphorimager use that.

Colin



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