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Nucleic Acids Res. 1996 Nov 1;24(21):4360-1.

A simple method for 3'-labeling of RNA.

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  • 1Department of Molecular Biology, Massachusetts General Hospital, Boston 02114, USA.


We describe a simple method for 3'-end labeling RNAs of known sequence. A short DNA template is designed to anneal to the 3'-end of the RNA, with a two nucleotide 5' overhang of 3'-TA-5', 3'-TG-5' or 3'-TC-5'. The Klenow fragment of DNA polymerase I can then cleanly and efficiently extend the 3'-end of the RNA by the incorporation of a single alpha-32P-labeled dATP residue. This method can be used to label one RNA in a mixture of RNAs, or to label 5'-blocked RNAs such as mRNA.

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