Analysis of off-target transcript levels. One potential off-target predicted with RNAhybrid was tested for smiRNA(CHS)-mediated effects on transcript abundance. RNA was extracted from different transgenic lines that highly over-expressed pri-smiRNA(CHS), and qRT-PCR experiments were performed to quantify the relative transcript levels of At1g40390. Data were normalized to wild type (wt) transcript levels. See also Figure 2B.