Solubilzation in formamide protects RNA from degradation

Abstract

Degradation by RNase is a major concern when handling RNA samples. The omnipresent enzyme (fingertips, dust etc.) is heat- stable and does not require cofactors for its enzymatic activity. This makes it difficult to inactivate RNase in RNA samples. Although there are several isolation methods yielding RNase-free RNA (1, 2), precautions have to be undertaken to avoid RNase contamination at the final step of isolation, i.e. solubilization of RNA. The common practice is to solubilize RNA in diethyl pyrocarbonate (DEPC)-treated water (1, 2). The solubilized RNA is susceptible, however, to accidental contamination with RNase during handling and storage of samples. In addition, long-term storage of water-solubilized RNA samples requires temperature as low as -70C to prevent degradation of RNA (1, 2).