Fingerprinting nonradioactive ribonucleic acid with the aid of polynucleotide phosphorylase.

Nucleic acids research ·Vol. 1 ·No. 1 ·1974-01-00 ·Pages 171-81

Szeto KS, Söll D

Abstract

We describe a method for obtaining radioactive fingerprints from nonradioactive ribonucleic acid. Fragments derived by T1 ribonuclease digestion of RNA are dephosphorylated with bacterial alkaline phosphatase. When these fragments are used as primers for the reaction of primer dependent polynucleotide phosphorylase with [alpha-(32)P]GDP in the presence of T1 ribonuclease the 3'-hydroxyl group of each fragment becomes phosphorylated. The degree of phosphorylation is reasonably uniform. The method has been applied to T1 ribonuclease digests of Escherichia coli tRNA(Met) (f); the oligonucleotides were further analyzed by spleen phosphodiesterase digestion. In a similar manner fingerprints of pancreatic ribonuclease digests of RNA can be obtained, when [alpha-(32)P]UDP, polynucleotide phosphorylase and pancreatic ribonuclease are used.

MeSH Terms

Alkaline Phosphatase Animals Electrophoresis, Gel, Two-Dimensional Escherichia coli/chemistry Phosphorus Radioisotopes Phosphorylation Polyribonucleotide Nucleotidyltransferase RNA/chemistry,isolation & purification RNA, Bacterial/chemistry,isolation & purification RNA, Transfer, Met/chemistry,isolation & purification Ribonucleases

Chemicals

Phosphorus Radioisotopes RNA, Bacterial RNA, Transfer, Met RNA Polyribonucleotide Nucleotidyltransferase Ribonucleases Alkaline Phosphatase

Authors & Affiliations

2 authors, click to expand affiliations / ORCID

Szeto K S

Söll D

References (10)

10 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/1.1.171

Published

1974-01-00

Pages

171-81

Language

English

Region

England

NLM ID

0411011

PMCID

PMC343333

Subset

External Links

PubMed source DOI: 10.1093/nar/1.1.171 PMC Full Text View journal details

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Fingerprinting nonradioactive ribonucleic acid with the aid of polynucleotide phosphorylase.

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