3'End labelling of RNA with 32P suitable for rapid gel sequencing.
A new general method of labelling the 2',3'-diol end of RNA with 32P has been devised suitable for gel sequencing. Poly(A) polymerase (E. coli) is incubated with the RNA and limiting amounts of alpha-32P-ATP. The mono-addition product is then cleaved with periodate and beta-eliminated with aniline, leaving the RNA terminally labelled with 3' 32P-phosphate. When applied to a model compound, tRNAPhe from E. coli, over 28 residues could be read from the 3' end.
D000255:Adenosine Triphosphate / Q000378:metabolism; D001483:Base Sequence; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D004926:Escherichia coli; D008722:Methods; D009690:Nucleic Acid Conformation; D010084:Oxidation-Reduction; D010649:Phenylalanine; D010761:Phosphorus Radioisotopes; D011062:Polynucleotide Adenylyltransferase / Q000378:metabolism; D012313:RNA; D012343:RNA, Transfer
D010761:Phosphorus Radioisotopes; D010649:Phenylalanine; D012313:RNA; D000255:Adenosine Triphosphate; D012343:RNA, Transfer; D011062:Polynucleotide Adenylyltransferase