3'End labelling of RNA with 32P suitable for rapid gel sequencing.

Nucleic acids research ·Vol. 5 ·No. 9 ·1978-09-00 ·Pages 3129-39

Winter G, Brownlee GG

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.

MeSH Terms

Adenosine Triphosphate/metabolism Base Sequence Electrophoresis, Polyacrylamide Gel/methods Escherichia coli Methods Nucleic Acid Conformation Oxidation-Reduction Phenylalanine Phosphorus Radioisotopes Polynucleotide Adenylyltransferase/metabolism RNA RNA, Transfer

Chemicals

Phosphorus Radioisotopes Phenylalanine RNA Adenosine Triphosphate RNA, Transfer Polynucleotide Adenylyltransferase

Authors & Affiliations

2 authors, click to expand affiliations / ORCID

Winter G

Brownlee G G

References (21)

21 references, click to expand

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Article Info

Journal

Nucleic acids research

Abbr.

Nucleic Acids Res

ISSN

0305-1048

DOI

10.1093/nar/5.9.3129

Published

1978-09-00

Pages

3129-39

Language

English

Region

England

NLM ID

0411011

PMCID

PMC342236

Subset

External Links

PubMed source DOI: 10.1093/nar/5.9.3129 PMC Full Text View journal details

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3'End labelling of RNA with 32P suitable for rapid gel sequencing.

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