Purification of RNA from cells and tissues by acid phenol-guanidinium thiocyanate-chloroform extraction

摘要

This protocol describes a single-step technique for the purification of RNA. Cells are homogenized in guanidinium thiocyanate and the RNA is purified from the lysate by extraction with phenol-chloroform at reduced pH(1). This method allows many samples to be processed simultaneously and quickly. The yield of total RNA depends on the tissue or cell source and is generally in the range of 4-7 mu g per ml starting tissue or 5-10 mu g per 10(6) cells. All reagents used in this protocol must be prepared with diethyl pyrocarbonate (DEPC)-treated H2O.