HELP!!! DNAse treating human adipose RNA

Jim Graham graham at biodec.wustl.edu
Thu Jul 17 10:54:18 EST 1997


Ivor,

Try "washing" the extracted RNA several times by resuspending in 
guanidiuium lysis buffer and alcohol reprecipitation. After two or three 
rounds, incubate an aliquot of the sample in your DNAse buffer at 37C 
for about 30 minutes and compare it on a fomraldehyde get to an 
untreated sample (eg. resuspended in GuThCy or loading buffer only). 
You need to establish that your RNA is stable in the DNase treatment.

(Store all RNAs as alcohol precipitated aliquots -by the way, and don't 
heat RNA over 70C, particularily if you have not chelated metal ions 
with EDTA first -and even if you have. :)

Jim
J. Graham PhD
Biology Department



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