End labeling of enzymatically decapped mRNA

End labeling of enzymatically decapped mRNA

A method is presented for rapid and efficient 5′ end label ing with 32 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...

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Veröffentlicht in:Nucleic acids research 1977-12, Vol.4 (12), p.4165-4174
Hauptverfasser: Efstratiadis, Argiris, Vournakis, John N., Donis-Keller, Helen, Chaconas, George, Dougall, Don K., Kafatos, Fotis C.
Format: Artikel
Sprache:eng
Schlagworte:
Adenosine Triphosphatases - metabolism
Alkaline Phosphatase
Diphosphates - metabolism
Isotope Labeling - methods
Kinetics
Phosphorus Radioisotopes
Plants - enzymology
RNA, Messenger
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Beschreibung
Zusammenfassung:A method is presented for rapid and efficient 5′ end label ing with 32 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 γ-32-P-ATP and T4 polynucleotide kinase.
ISSN:0305-1048
1362-4962
DOI:10.1093/nar/4.12.4165