title: Synthesis and labeling of RNA in vitro.
abstract: This unit discusses several methods for generating large amounts of uniformly labeled, end-labeled, and site-specifically labeled RNAs in vitro. The methods involve a number of experimental procedures, including RNA transcription, 5' dephosphorylation and rephosphorylation, 3' terminal nucleotide addition (via ligation), site-specific RNase H cleavage directed by 2'-O-methyl RNA-DNA chimeras, and 2-piece splint ligation. The applications of these RNA radiolabeling approaches are also discussed.
authors.affiliation: Process Science Downstream, Bristol-Myers Squibb Company, East Syracuse, New York, USA.
authors.forename: Chao
authors.initials: C
authors.lastname: Huang
authors.forename: Yi-Tao
authors.initials: YT
authors.lastname: Yu
mesh_terms: D066298:In Vitro Techniques; D012313:RNA / Q000138:chemical synthesis / Q000737:chemistry / Q000235:genetics; D011868:Radioisotopes; D016914:Ribonuclease H / Q000737:chemistry; D013194:Staining and Labeling / Q000379:methods*; D013379:Substrate Specificity; D014158:Transcription, Genetic
chemical_list: D011868:Radioisotopes; D012313:RNA; D016914:Ribonuclease H
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