Enrichment of biologically active U1 small nuclear RNAs by ion-exchange high-performance liquid chromatography

The use of ion-exchange high-performance liquid chromatography in conjunction with preparative electrophoresis to facilitate the purification of biologically active snRNAs is described. Separation of total nuclear RNA from a Bombyx mori cell line was done with a Bio-Rad MA7 plasmid column in a HRLC 500 system. Individual fractions were subjected to electrophoresis through 14% polyacrylamide gels for identification. High levels of U1 RNA were confirmed by Northern analysis with a human U1 probe. Biological activity of RNAs from the column was demonstrated by their ability to incorporate 32P-AMP at the 3' end. Ion-exchange chromatography provides a rapid, automated method for purifying large amounts of RNAs that can then be utilized in further studies.