Isolation and Purification of RNA

Abstract

RNA extraction and handling are technically more challenging than DNA manipulation due to the presence of a reactive 2'-OH on RNA ribose residues and to the ubiquitous and stable nature of RNases.1, 2 Consequently, signi cant measures should be taken to preserve RNA integrity throughout puri cation. Rapid inactivation of RNases released during cell lysis is critical for the isolation of high-quality RNA. All RNA extraction buffers, therefore, contain powerful inactivating denaturants such as guanidinium salts or detergents. Successful RNA extraction relies on good laboratory practice and RNase-free technique. Since airborne bacteria and the skin (e.g., hands) are also sources of RNases, all solutions and labware used in RNA isolation should be treated to remove RNases, and gloves must be worn at all times. We recommend the following precautions for RNA work.