Figure 1.

HyCCAPP is a multistep process that uses sequence-specific hybridization to capture DNA loci of interest from formaldehyde cross-linked cells. The overall procedure involves (1) cross-linking cells with formaldehyde followed by cell lysis, (2) hybridization capture from sonicated and RNaseA-treated lysate using desthiobiotin oligonucleotide capture probes and streptavidin-coated magnetic beads, (3) mass spectrometric analysis of captured proteins using LC–MS/MS, and (4) bioinformatic analysis of mass spectrometric data to determine the proteins enriched at the captured DNA locus of interest relative to non-enriched yeast lysate.