RNA quantification & Northerns

SSD4 at psuvm.psu.edu SSD4 at psuvm.psu.edu
Wed Apr 14 07:08:26 EST 1993


Does anyone out there have the secret for obtaining repeatability in
loading northerns?  Currently I am using A260 to quantify the RNA in my
samples and then determining the volume required for 10 ug.  After drying
in a speed vac I dissolve the RNA in 2.2 ul of 3X MOPS/EDTA before adding
4.8 ul of formaldehyde/formamide mixture and heating to denature the RNA.
Regardless of the time and attention spent in sample preparation I
invariably end up with inequal loading (based on staining with ethidium).
Is there a better method than absorbance @ 260 for quantifying RNA or is
there a possible problem in my sample prep.

Fishing for answers / suggestions

Shawn S. Donkin



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