Synthesis and annealing properties of RNAs containing randomized nucleotide positions. (A) Sequences of the siRNDs 1–6. Watson–Crick base pairs are indicated with a dash. Randomized vicinal positions are indicated with a dot. (B) Deep-sequencing analysis of siRND3 antisense strand shows equal base distribution at nearly all randomized positions. Consensus sequence was created using WebLogo3. (C) Native polyacrylamide gel electrophoresis (PAGE) of siRNDs. Lanes 1,2, and 10 are loaded with single-stranded RNAs from siRND1 and siRND4 and serve as references; lane 9 contains siCon; lanes 3–8 contain siRNDs 1–6. (D) Melting curve analysis of siRNDs. The graph shows the first derivative of melting curves. TMs listed in the table represent the mean of triplicate measurements ± SD.
