Purification and characterization of transcribed RNAs using gel filtration chromatography
- 13 December 2007
- journal article
- Published by Springer Nature in Nature Protocols
- Vol. 2 (12) , 3270-3277
- https://doi.org/10.1038/nprot.2007.480
Abstract
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.Keywords
This publication has 29 references indexed in Scilit:
- Preparation of large RNA oligonucleotides with complementary isotope-labeled segments for NMR structural studiesNature Protocols, 2007
- Viral dsRNA Inhibitors Prevent Self-association and Autophosphorylation of PKRJournal of Molecular Biology, 2007
- Fluorescent Pyrimidine Ribonucleotide: Synthesis, Enzymatic Incorporation, and UtilizationJournal of the American Chemical Society, 2007
- Rapid purification of RNAs using fast performance liquid chromatography (FPLC)RNA, 2006
- Specific Recognition of HIV TAR RNA by the dsRNA Binding Domains (dsRBD1–dsRBD2) of PKRJournal of Molecular Biology, 2006
- Uncoupling of RNA Binding and PKR Kinase Activation by Viral Inhibitor RNAsJournal of Molecular Biology, 2006
- In vitro Transcription and Purification of RNAs of Different SizeChimia, 2005
- Structure of HCV IRES domain II determined by NMRNature Structural & Molecular Biology, 2003
- NMR Study of 100 kDa HCV IRES RNA Using Segmental Isotope LabelingJournal of the American Chemical Society, 2002
- Crystallization of RNA-protein complexes I. Methods for the large-scale preparation of RNA suitable for crystallographic studiesJournal of Molecular Biology, 1995