Native RNA purification by gel filtration chromatography.
In vitro transcription of RNA from DNA templates by T7 RNA polymerase allows for the generation of large quantities of RNA suitable for many downstream applications. The resulting RNA can be purified by a number of methodologies. Herein, we describe the native isolation of RNA molecules by FPLC purification using Superdex 75 or 200 gel filtration columns. This approach can be extended to purify biologically interesting RNA complexes such as RNA-protein complexes that have been generated from either synthetic or in vitro transcribed RNAs and recombinant proteins.
Department of Chemistry, University of Manitoba, Winnipeg, MB, Canada.
Evan P
EP
Booy
Hui
H
Meng
Sean A
SA
McKenna
D002021:Buffers; D002498:Centrifugation; D002850:Chromatography, Gel / Q000379:methods*; D002851:Chromatography, High Pressure Liquid; D021141:Nucleic Acid Amplification Techniques; D012313:RNA / Q000096:biosynthesis / Q000302:isolation & purification*; D014158:Transcription, Genetic
D002021:Buffers; D012313:RNA