A simple method for 3'-labeling of RNA.
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.
Huang
Z
Z
Department of Molecular Biology, Massachusetts General Hospital, Boston 02114, USA.
Szostak
J W
JW
D000345:Affinity Labels; D004247:DNA / Q000378:metabolism; D004256:DNA Polymerase I / Q000378:metabolism; D003838:Deoxyadenine Nucleotides / Q000378:metabolism; D005821:Genetic Techniques; D006801:Humans; D010761:Phosphorus Radioisotopes; D012313:RNA / Q000378:metabolism; D013698:Templates, Genetic
D000345:Affinity Labels; D003838:Deoxyadenine Nucleotides; D010761:Phosphorus Radioisotopes; D012313:RNA; D004247:DNA; D004256:DNA Polymerase I; C026600:2'-deoxyadenosine triphosphate