Centromere Specificaton ChIP-chip arrays

Cenromere Specificaton and Kinetochore Function (Lieb project, Desai subgroup)

General Description

Synchronized C. elegans early embryos from strain N2 were treated with the cross-linking reagent formaldehyde. Sonicated chromatin was prepared and immunoprecipitated with an affinity-purified polyclonal antibody that recognizes the centromere-binding protein HCP-3 (CENP-A). After whole genome amplification, NimbleGen genomic tiling microarrays were used in two-color hybridization experiments to compare the signal from the input DNA versus the fragments pulled-down in the ChIP. Normalized log2(ratios) are shown in this track.

Protocols

  1. Growth and isolation: Worm embryo extraction:JL:RG1, Worm embryo growth and harvest:JL:RG1
  2. Sample preparation: ChIP-chip label hyb nimblegen:JL:1, ChIP-chip scanning nimblegen:JL:2, Worm LM-PCR Amplification for ChIP-chip:JL:RG1, Worm Chromatin Immunoprecipitation:JL:RG1, ChIP-chip scanning nimblegen:JL:1, ChIP-chip label hyb nimblegen:JL:2
  3. Other Protocols: ChIP-chip normalization standard nimblegen:JL:1

Experimental Reagents

    Growth Conditions:
  1. Antibodies: OD00079_HCP3, OD00001_HCP3
  2. Arrays: 080922_MODENCODE_CE_CHIP_HX1, NimbleGen DesignName2006-07-18 C elegans ChIP03, NimbleGen DesignName2006-07-18 C elegans ChIP02, NimbleGen DesignName2006-07-18 C elegans ChIP01

Sample Details

  1. Animals/Lines: Mixed Stage Embryos


Release Date: 2009-10-15