3'End labelling of RNA with 32P suitable for rapid gel sequencing
- PMID: 360168
- PMCID: PMC342236
- DOI: 10.1093/nar/5.9.3129
3'End labelling of RNA with 32P suitable for rapid gel sequencing
Abstract
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.
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