Purification and characterization of transcribed RNAs using gel filtration chromatography.
RNA synthesis using in vitro transcription by phage T7 RNA polymerase allows preparation of milligram quantities of RNA for biochemical, biophysical and structural investigations. Previous purification approaches relied on gel electrophoretic or gravity-flow chromatography methods. We present here a protocol for the in vitro transcription of RNAs and subsequent purification using fast-performance liquid chromatography. This protocol greatly facilitates production of RNA in a single day from transcription to purification.
McKenna
Sean A
SA
Department of Structural Biology, Stanford University School of Medicine, Stanford, California, USA.
Kim
Insil
I
Puglisi
Elisabetta Viani
EV
Lindhout
Darrin A
DA
Aitken
Colin Echeverría
CE
Marshall
R Andrew
RA
Puglisi
Joseph D
JD
D002850:Chromatography, Gel; D010957:Plasmids; D012313:RNA / Q000032:analysis*; D016337:RNA, Catalytic; D045162:Thiazolidinediones; D014158:Transcription, Genetic
D016337:RNA, Catalytic; D045162:Thiazolidinediones; C427204:hammerhead ribozyme; C114240:DN 108; D012313:RNA