Project description:ChIP-chip was performed using chromatin isolated from formaldehyde crosslinked, staged embryos, and affinity purified antibodies against HB and MED. Affinity purification of the antibodies was performed with E.coli expressed His-tag fusions of the N-terminal portion of the full length protein (designated as 1, e.g. anti-HB 1 ), or the C-terminal portion of the protein (designated as 2, e.g. anti-HB 2, and anti_MED 2). The chromatin immunoprecitation and hybridization for each antibody were carried out in duplicates, and in each chromatin immunoprecitation and hybridization series, two mock IP controls and two input DNA controls were also generated. The microarray used in this study is the Affymetrix Drosophila genomic DNA tiling array, which has about 3 million oligo pairs (perfect match-mismatch), covering the whole euchromatic sequences of the fly genome at a resolution of about 35 bp.
Project description:RIN-associated mRNAs were identified in early Drosophila embryos by RNA co-immunoprecipitation of the endogenous protein using a synthetic antibody (anti-RIN D072), followed by microarray analysis (RIP-Chip).
