Hydrolysis of single-stranded RNA in aqueous solutions—effect on quantitative hybridizations

Abstract

Single stranded RNA is non-enzymatically hydrolysed in aqueous solutions at neutral pH at elevated temperatures. This hydrolysis causes practical problems in different hybridization procedures. Synthetic cRNA transcribed from the human p53 oncogene was found to be partially destroyed after 6 hours and completely destroyed after overnight incubation at 60 °C. At lower temperatures the cRNA was preserved intact in spite of overnight incubation, but at higher temperatures (80 °C) the degradation occurred in less than 2 hours. The effect of the hydrolysis on hybridization results was studied by measuring in solution the hybridization kinetics of the cRNAs of another human oncogene, N- myc. It is obvious that conditions generally used for DNA hybridizations cannot be utilized for RNA in quantitative studies.