ChIP-chip: Histone Modifications in Pupae

Pupae H3K27Ac ChIP-chip (White project)

Details

Nuclei were isolated from staged Drosophila (y; bw cn sp) pupae, and cross-linked with formaldehyde. After lysis and sonication the chromatin was immunoprecipitated with affinity-purified antibodies recognizing histone H3 acetylated on lysine 27. The DNA was amplified, labeled, and hybridized to Agilent microarrays (mock IP DNA was labelled in parallel and hybridized simultaneously to the same chip). A Joint Binding Deconvolution algorithm was used to predict binding peaks in normalized data merged from three experimental replicates.

Series Description

The White Lab is aiming to map the association of all the Transcription Factors (TF) and DNA associated proteins on the genome of Drosophila melanogaster. The main technique that will be used for this purpose is chromatin immunoprecipitation-on-chip (ChIP-on-chip) utilizing whole-genome tiling arrays. The data generated by ChIP-chip experiments consist basically of a plot of signal intensity across the genome. The highest signals correspond to positions in the genome occupied by the tested TF.

Protocols

  1. Growth and isolation: Pupae collection histone timecourse:KW:1
  2. Sample preparation: White_Lab/IP, Whole_Genome_Amplification, Agilent 244K ChIP Labeling:KW:1, Array Hybridization for ChIP-chip:KW:1, Array Scanning
  3. Other Protocols: Cross-linking, Agilent ChIP data processing:KW:1, Agilent ChIP peak finding:KW:1

Experimental Reagents

  1. Antibodies: H3K27Ac, No Antibody Control
  2. Arrays: Agilent Drosophila tiling array 1 of 3, Agilent Drosophila tiling array 2 of 3, Agilent Drosophila tiling array 3 of 3

Sample Details

  1. Animals/Lines: Pupae, Drosophila melanogaster,


Release Date: 2010-01-12