Figure 2.

Optimization of assay conditions (I). Optimization of (A) incubation temperature, (B) pH of ligation buffer, (C) pH of annealing buffer. (D) Assay performance with dithiothreitol (DTT) versus β-mercaptoethanol (2-ME) as reducing agent. For (B), (C) and (D), parallel assays without REL1 were carried out as controls. Except for the parameters tested, all experiments were carried out as described in ‘Materials and Methods’ section under ‘Initial RNA annealing and ligation conditions’, and means and standard deviations for three replicates are shown.