A siRNA screen was conducted with a small siRNA library against PGL3 luciferase. a, Depiction of the small siRNA library against PGL3 luciferase. The target sequence for the siRNA against PGL3 siRNA that was tested in Figure 3b is shown. The bases in bold were randomized by synthesizing an oligo with random bases in place of CTC and cloned into the pHippy vector. b, Screening of the siRNA library. 12 pools of 10 bacterial colonies from the random siRNA library against PGL3 luciferase were screened for their ability to inhibit PGL3 luciferase activity as described in the legend to Figure 3b. c, The sequences of the siRNA single clones, which did or did not inhibit PGL3 luciferase. Pools 8 and 11 were broken down into individual clones and re-screened for inhibitory activity. Pool 8 contained 1 positive clone and Pool 11 contained 2 positive clones. The sequences of all three positive clones is shown on the left and corresponds to the original PGL3 siRNA. The sequences of 10 negative clones are shown on the right.
Kaykas and Moon BMC Cell Biology 2004 5:16 doi:10.1186/1471-2121-5-16