title: In vitro transcription from plasmid or PCR-amplified DNA.
abstract: This protocol describes the synthesis and purification of RNAs using plasmid DNA or PCR-amplified DNA as a template. This procedure should give NTP-free, full-length RNA for all sizes of RNA. This protocol is derived from Milligan and Uhlenbeck, the classic paper on T7 transcription reactions, with modifications.
authors.affiliation: Howard Hughes Medical Institute, Johns Hopkins University School of Medicine, Baltimore, MD, USA; Department of Molecular Biology and Genetics, The Johns Hopkins University School of Medicine, Baltimore, MD, USA.
authors.forename: Julie L
authors.initials: JL
authors.lastname: Brunelle
authors.forename: Rachel
authors.initials: R
authors.lastname: Green
mesh_terms: D017123:Bacteriophage T7 / Q000235:genetics*; D004247:DNA / Q000235:genetics* / Q000302:isolation & purification; D019800:Phenol / Q000737:chemistry; D010957:Plasmids / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012313:RNA / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification; D013698:Templates, Genetic; D014158:Transcription, Genetic
chemical_list: D019800:Phenol; D012313:RNA; D004247:DNA
keywords: Column purification; In vitro transcription reaction; PCR-amplified DNA; Phenol extract DNA Template; RNA polymerase