End labeling of enzymatically decapped mRNA

A Efstratiadis; J N Vournakis; H Donis-Keller; G Chaconas; D K Dougall; F C Kafatos

doi:10.1093/nar/4.12.4165

End labeling of enzymatically decapped mRNA

Nucleic Acids Res. 1977 Dec;4(12):4165-74. doi: 10.1093/nar/4.12.4165.

Authors

A Efstratiadis, J N Vournakis, H Donis-Keller, G Chaconas, D K Dougall, F C Kafatos

Abstract

A method is presented for rapid and efficient 5' end labeling with 32P of capped mRNAs, by a series of three enzymatic reactions: the blocking nucleotide of the cap structure is removed by tobacco acid pyrophosphatase, and after dephosphorylation with alkaline phosphatase the 5' end is labeled with gamma-32-P-ATP and T4 polynucleotide kinase.

Publication types

Research Support, U.S. Gov't, Non-P.H.S.
Research Support, U.S. Gov't, P.H.S.

MeSH terms

Adenosine Triphosphatases / metabolism
Alkaline Phosphatase
Diphosphates / metabolism
Isotope Labeling / methods
Kinetics
Phosphorus Radioisotopes
Plants / enzymology
RNA, Messenger*

Substances

Diphosphates
Phosphorus Radioisotopes
RNA, Messenger
Alkaline Phosphatase
Adenosine Triphosphatases