Characterization and use of the potent ribonuclease inhibitor aurintricarboxylic acid for the isolation of RNA from animal tissues.
Inclusion of aurintricarboxylic acid (ATA) in extraction buffers for the isolation of RNA from animal tissues resulted in high yields (0.5-2.0 mg/g of tissue) of undegraded material as judged by agarose-gel-electrophoretic analyses and Northern-blotting experiments. However, ATA bound to nucleic acids, forming stable complexes, and so we have established methods for spectrophotometric quantification of RNA in these coloured complexes, and for easy removal of sufficient ATA to leave RNA in a consistently hybridizable condition at the end of a purification. The use and subsequent removal of ATA was straightforward and gave satisfactory results for all rat tissues tested.
Skidmore
A F
AF
Department of Biochemistry, University of Sussex, Falmer, Brighton, U.K.
Beebee
T J
TJ
D000818:Animals; D001312:Aurintricarboxylic Acid / Q000032:analysis; D015152:Blotting, Northern; D003509:Cyclohexanecarboxylic Acids / Q000032:analysis; D004247:DNA / Q000378:metabolism; D004587:Electrophoresis, Agar Gel; D007202:Indicators and Reagents; D009693:Nucleic Acid Hybridization; D012313:RNA / Q000302:isolation & purification*; D051381:Rats; D012260:Ribonucleases / Q000037:antagonists & inhibitors*; D012474:Salmon; D013056:Spectrophotometry, Ultraviolet
D003509:Cyclohexanecarboxylic Acids; D007202:Indicators and Reagents; D001312:Aurintricarboxylic Acid; D012313:RNA; D004247:DNA; D012260:Ribonucleases