Figure 3

Schematic of the HaloCHIP-chip microarray experiment design. HaloCHIP DNA (10-50 ng) obtained for both the experimental HaloTag-CREB and untransfected control sample was purified and amplified to a concentration of 1-10 μg using the whole genome amplification (WGA) method (Sigma) [46]. The HaloTag-CREB amplified sample was labelled with Cy5 (green) and the untransfected control sample with Cy3 (red), then hybridized to a custom DNA oligo microarray manufactured by Roche NimbleGen. The oligo array was designed to cover on average a 1.8 kb region of 27,661 human promoter regions that contain 33,255 TSS predicted by SwitchGear Genomics. To obtain coverage of each promoter, an average of fourteen 50mer single stranded DNA probes, shown in purple, per promoter were used, with an average spacing of 131 bp per probe.