End labeling of enzymatically decapped mRNA.
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.
D000251:Adenosine Triphosphatases / Q000378:metabolism; D000469:Alkaline Phosphatase; D011756:Diphosphates / Q000378:metabolism; D007553:Isotope Labeling / Q000379:methods; D007700:Kinetics; D010761:Phosphorus Radioisotopes; D010944:Plants / Q000201:enzymology; D012333:RNA, Messenger
D011756:Diphosphates; D010761:Phosphorus Radioisotopes; D012333:RNA, Messenger; D000469:Alkaline Phosphatase; D000251:Adenosine Triphosphatases