title: Enrichment of biologically active U1 small nuclear RNAs by ion-exchange high-performance liquid chromatography.
abstract: 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.
authors.affiliation: Department of Biological Sciences, Florida International University, Miami 33199.
authors.forename: V
authors.initials: V
authors.lastname: Leff
authors.forename: J P
authors.initials: JP
authors.lastname: Gao
authors.forename: L R
authors.initials: LR
authors.lastname: Vega
authors.forename: R J
authors.initials: RJ
authors.lastname: Herrera
mesh_terms: D000249:Adenosine Monophosphate / Q000378:metabolism; D000818:Animals; D015152:Blotting, Northern; D012831:Bombyx; D002460:Cell Line; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D002852:Chromatography, Ion Exchange / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D010053:Ovary / Q000737:chemistry / Q000166:cytology / Q000378:metabolism; D010758:Phosphorus / Q000378:metabolism; D010957:Plasmids; D015347:RNA Probes; D012342:RNA, Small Nuclear / Q000302:isolation & purification* / Q000378:metabolism
chemical_list: D015347:RNA Probes; D012342:RNA, Small Nuclear; D010758:Phosphorus; D000249:Adenosine Monophosphate