H4 Histone modifications in S2-DRSC cells

H4 Histone modifications in S2-DRSC cells (Karpen project, Kuroda subgroup)

General Description

We aim to determe the locations of 125 chromosomal proteins and histone modifications across the Drosophila melanogaster genome. The proteins and modifications under study are involved in basic chromosomal functions such as DNA replication, gene expression, gene silencing, and inheritance. We will perform Chromatin ImmunoPrecipitation (ChIP) using genomic tiling arrays. We will initially assay localizations using chromatin from three cell lines and two embryonic stages, and will then extend the analysis of a subset of proteins to four additional animal tissues/stages.

Protocols

  1. Growth and isolation: Chromatin prep from cultured cells, Crosslinking of cultured cells
  2. Sample preparation: Hybridization to Affy arrays, WGA, ChIP, Array Scanning Protocol
  3. Data Analysis: Smoothed M-value enrichment profiles, M-value normalization, Smoothed M-value enrichment profiles, Regions of significant enrichment

Experimental Reagents

  1. Antibodies: Ab:H4K20me:GK:1, H4AcTetra, H4K16ac(L), H4K8ac, H4K5ac, H4K16ac(M)
  2. Arrays: Affymetrix Drosophila Tiling Arrays v2.0R

Sample Details

  1. Animals/Lines: S2-DRSC

  1. Other Protocols: Regions of significant enrichment


Release Date: 2009-10-11