Skip to main content
. 2012 Jul 23;40(20):e157. doi: 10.1093/nar/gks698

Figure 4.

Figure 4.

The detection of the Am1164 2'-O-methylation in the 25S rRNA of S. pombe by RTL-P. (A) The potential base-pairing interaction between the snR61 snoRNA and its target region in the 25S rRNA of S. pombe. (B) A schematic diagram of the RT primer design. (C) An analysis of Am1164 by the traditional primer extension method in wild type (Wt) and mutant (△snR61) S. pombe. The primer extension was performed in the presence of increasing dNTP concentrations (4 μM and 0.15 mM). A subset of the rRNA sequence is indicated on the left. The position of the reverse transcriptase stop site is indicated by an arrow, and the methylated site is marked with an asterisk. T/G/C/A = the rDNA sequence ladders. (D and E) The detection of Am1164 by RTL-P at varying dNTP concentrations with 20 ng (D) and 2 ng total RNA (E) in each RT reaction. All assays were independently carried out at least three times, and the representative results of a single experiment are shown. The ratio of PCR signal intensity is shown beneath each lane.