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.
Leff
V
V
Department of Biological Sciences, Florida International University, Miami 33199.
Gao
J P
JP
Vega
L R
LR
Herrera
R J
RJ
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
D015347:RNA Probes; D012342:RNA, Small Nuclear; D010758:Phosphorus; D000249:Adenosine Monophosphate