Application of the high-resolution northern blotting in monitoring RNA reagents expressed in cells. A) Processing of antisense-sense and sense-antisense shRNAs (L-shSCA3 and R-shSCA3, respectively). The 5' and 3' strands of each shRNA were analyzed by detection with probes complementary to either siRNA strand, as indicated in the figure. One probe detected L-5' and R-3' strands (dotted line) while the other probe detected L-3' and R-5' strands (dashed line). B) ssRNA CUG9 and shRNA CAG/CUG7 transcribed from vectors, having either 4T or 6T at their termination sites. M denotes size markers; M1 denotes end-labeled 17, 19, 21, 23, and 25-nt synthetic RNA oligonucleotides and M2 denotes RNA Low Molecular Weight Marker (USB). Quantitative representations of siRNA length variants are shown in the bottom panel, using peaks obtained from phosphorimaging analyses.
Koscianska et al. BMC Molecular Biology 2011 12:14 doi:10.1186/1471-2199-12-14