title: 3'-End labeling of RNA with yeast Poly(A) polymerase and 3'-deoxyadenosine 5'-[α-32P]triphosphate.
abstract: This protocol is used to label RNA molecules (in vitro-synthesized or in vivo-purified RNA molecules) that have free 3'-hydroxyl termini. The reaction is performed in 10 min using yeast poly(A) polymerase and 3'-deoxyadenosine 5'-[α-(32)P]triphosphate (cordycepin 5'-[α-(32)P]triphosphate), a chain-terminating nucleotide. At the end of the procedure, the reaction is desalted by gel filtration to remove any unincorporated nucleotides.
authors.forename: Donald C
authors.initials: DC
authors.lastname: Rio
mesh_terms: D003838:Deoxyadenine Nucleotides / Q000378:metabolism*; D008967:Molecular Biology / Q000379:methods*; D010761:Phosphorus Radioisotopes / Q000378:metabolism*; D011062:Polynucleotide Adenylyltransferase / Q000378:metabolism*; D012313:RNA / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000201:enzymology*; D013194:Staining and Labeling / Q000379:methods*; D013997:Time Factors
chemical_list: D003838:Deoxyadenine Nucleotides; D010761:Phosphorus Radioisotopes; C011746:3'-deoxyadenosine 5'-triphosphate; D012313:RNA; D011062:Polynucleotide Adenylyltransferase