Chip-Seq (Chromatin Silencers Antibodies)

Chromatin Silencers ChIP-Seq (White project, White subgroup)

Nuclei were isolated from staged Drosophila (y; cn bw sp) embryos, 0- 12 hr after egg laying, and cross-linked with formaldehyde. After lysis and sonication the chromatin was immunoprecipitated with affinity-purified antibodies recognizing histone deacetylases. The DNA was amplified, labeled, and hybridized to Input and ChIP'd DNA was single-end sequenced in two lanes each on the Illumina Genome Analyzer, aligned with Eland. Density ratios were calculated with the R SPP package. PeakSeq software was used to call peaks.

General 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.
ChIP-seq, meaning direct sequencing of the ChIP sample on the Solexa platform, has also been used to validate some datasets and will become- as the project advances- the main production platform.

To support ChIP-chip and ChIP-seq datasets, and in order to link association of factors to the DNA with regulation of transcription, several RNA-seq (direct sequencing of the transcriptome on the Solexa platform) datasets will be genrated for the time-points under scrutiny.

Protocols

  1. Growth and isolation: Embryonic collection Stage 0-12 hours AEL:KW:1

Experimental Reagents

  1. Antibodies: HDAC4a-492, HDAC3-499, HDAC3-498, HDAC4a-493, No Antibody Control, HDAC11-495, HDAC1-500, HDAC11-494, HDAC6-497, HDAC6-496, HDAC1-501 Sin3A-RC

Sample Details

  1. Animals/Lines: y[1]; Gr22b[1] Gr22d[1] cn[1] CG33964[R4.2] bw[1] sp[1]; LysC[1] MstProx[1] GstD5[1] Rh6[1], Embryo 0-12h
  2. External Links: GSM499635, GSM499633, GSM499642, GSM499640, GSM499634, GSM499639, GSM499636, GSM499634, GSM499638, GSM499637, GSM499643, GSM499641 GSM569791, GSM569792

Release Date: 2010-01-25