Journal: Nucleic Acids Research
Article Title: HEN1 recognizes 21-24 nt small RNA duplexes and deposits a methyl group onto the 2? OH of the 3? terminal nucleotide
Figure Lengend Snippet: Effect of various modifications on the 3′ terminal nucleotide of a small RNA on T4 RNA ligase- and yeast PAP-catalyzed reactions. ( a ) T4 RNA ligase-mediated ligation of various miR173 forms to an RNA linker. ( b ) Activity of yeast PAP on various forms of miR173 in the presence of 2 pmol [α- 32 P]-ATP. The ladders or smears represent products of PAP-catalyzed reaction. ( c ) Activity of yeast PAP on various forms of miR173 in the presence of 10 pmol [α- 32 P]-ATP.
Article Snippet: Ligation of the miR173 RNAs to an RNA linker (5′-pUAUGAAGCC), in which the 3′ OH is blocked by a C-3 spacer (CH2 CH2 CH2 OH), was performed at 16°C for 16 h in a 10 µl mixture containing 20 U T4 RNA ligase (Amersham Pharmacia), 1 ng of dephosphorylated miR173 and 1 µg RNA linker.
Techniques: Ligation, Activity Assay