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

Molecular and Cellular Probes
J Tenhunen

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 degrees C. At lower temperatures the cRNA was preserved intact in spite of overnight incubation, but at higher temperatures (80 degrees 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.

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Citations

Jul 19, 2012·World Journal of Microbiology & Biotechnology·Marimuthu CitartanThean-Hock Tang
Sep 1, 1991·Journal of Virological Methods·L RouhiainenH Söderlund
Oct 2, 2007·Nucleic Acids Research·Igor D VilfanNynke H Dekker
Jun 1, 1992·Molecular and Cellular Probes·E KagaS Uesugi
Dec 1, 1990·Genetic Analysis, Techniques and Applications·J TenhunenH Söderlund

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