CATCH-IT data, Aha labeling timecourse

Nucleosome Turnover in S2 cells: Kinetic Time Course

D. melanogaster S2 tissue culture cells were metabolically labeled with the methionine analog, azidohomoalanine (Aha). Isolated nuclei were biotinylated using biotin alkyne, and digested with micrococcal nuclease to yield biotin-labeled mononucleosomes. The soluble chromatin fraction (the "Input") was incubated with Streptavidin beads to isolate nucleosomes containing newly synthesized histone subunits. After extensive washing, genomic DNA (the "Pulldown") was extracted from the beads and amplified. Labeled Input and Pulldown DNA was used in two-color hybridization experiments on NimbleGen genomic tiling microarrays. Cells were labeled for 20, 40 or 60 minutes; the median log2 signal from two replicates is shown in this track.

Additional details can be found in: Genome-wide kinetics of nucleosome turnover determined by metabolic labeling of histones. Science 328:1161-4, (2010) by Deal RB, Henikoff JG, & Henikoff S. PMID: 20508129. The complete publicly released data set is available at GEO: GSE19788