Fingerprinting nonradioactive ribonucleic acid with the aid of polynucleotide phosphorylase

We describe a method for obtaining radioactive fingerprints from non-radioactive ribonucleic acid. Fragments derived by Tl ribonuclease digestion of RNA are dephosphorylated with bacterial alkaline phosphatase. When these fragments are used as primers for the reaction of primer dependent poly-nucleo...

Full description

Saved in:
Bibliographic Details
Published in:Nucleic acids research 1974-01, Vol.1 (1), p.171-182
Main Authors: Szeto, Kwok S., Söll, Dieter
Format: Article
Language:English
Subjects:
Alkaline Phosphatase
Animals
Electrophoresis, Gel, Two-Dimensional
Escherichia coli
Escherichia coli - chemistry
Phosphorus Radioisotopes
Phosphorylation
Polyribonucleotide Nucleotidyltransferase
Ribonucleases
RNA - chemistry
RNA - isolation & purification
RNA, Bacterial - chemistry
RNA, Bacterial - isolation & purification
RNA, Transfer, Met - chemistry
RNA, Transfer, Met - isolation & purification
Citations: Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:We describe a method for obtaining radioactive fingerprints from non-radioactive ribonucleic acid. Fragments derived by Tl ribonuclease digestion of RNA are dephosphorylated with bacterial alkaline phosphatase. When these fragments are used as primers for the reaction of primer dependent poly-nucleotide phosphorylase with [α-32P]GDP in the presence of Tl ribonuclease the 3′-hydroxyl group of each fragment becomes phosphorylated. The degree of phosphorylation is reasonably uniform. The method has been applied to Tl ribonuclease digests of Eecherichia coli tRNA f Met ; the oligonucleotides were further analyzed by spleen phosphodiesterase digestion. In a similar manner fingerprints of pancreatic ribonuclease digests of RNA can be obtained, when [α-32P]UDP, polynucleotide phosphorylase and pancreatic ribonuclease are used.
ISSN:0305-1048
1362-4962
DOI:10.1093/nar/1.1.171