Figure 1.

Two-step ligation of a dsRNA adaptor to blunt-ended dsRNA. (A) Illustration of the ligation method. The dsRNA substrate (red) was FAM-conjugated for easy detection in gels. In Step I, T4 RNA ligase attaches an adenine hexamer oligomer (green) to the dsRNA substrate producing an overhanging 5′ terminus (sticky terminus). In Step II, the sticky-ended dsRNA is phosphorylated by T4 polynucleotide kinase and joined to a dsRNA adaptor, which contains a complementary overhang (blue), using T4 DNA ligase. (B) Native and (C) denaturing PAGE analyses of the first and the second steps. Each band is indicated with a corresponding illustration.