title: Large scale purification of RNA nanoparticles by preparative ultracentrifugation.
abstract: Purification of large quantities of supramolecular RNA complexes is of paramount importance due to the large quantities of RNA needed and the purity requirements for in vitro and in vivo assays. Purification is generally carried out by liquid chromatography (HPLC), polyacrylamide gel electrophoresis (PAGE), or agarose gel electrophoresis (AGE). Here, we describe an efficient method for the large-scale purification of RNA prepared by in vitro transcription using T7 RNA polymerase by cesium chloride (CsCl) equilibrium density gradient ultracentrifugation and the large-scale purification of RNA nanoparticles by sucrose gradient rate-zonal ultracentrifugation or cushioned sucrose gradient rate-zonal ultracentrifugation.
authors.affiliation: Nanobiotechnology Center, Markey Cancer Center, Department of Pharmaceutical Sciences, University of Kentucky, 789 S. Limestone Avenue, Room #565, Lexington, KY, 40536-0596, USA.
authors.forename: Daniel L
authors.initials: DL
authors.lastname: Jasinski
authors.forename: Chad T
authors.initials: CT
authors.lastname: Schwartz
authors.forename: Farzin
authors.initials: F
authors.lastname: Haque
authors.forename: Peixuan
authors.initials: P
authors.lastname: Guo
mesh_terms: D002499:Centrifugation, Density Gradient / Q000379:methods*; D002586:Cesium / Q000737:chemistry; D002712:Chlorides / Q000737:chemistry; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D012321:DNA-Directed RNA Polymerases / Q000737:chemistry; D053758:Nanoparticles / Q000737:chemistry*; D012313:RNA / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification*; D014461:Ultracentrifugation / Q000379:methods*; D014764:Viral Proteins / Q000737:chemistry
chemical_list: D002712:Chlorides; D014764:Viral Proteins; D002586:Cesium; D012313:RNA; C062210:bacteriophage T7 RNA polymerase; D012321:DNA-Directed RNA Polymerases; C028019:cesium chloride