title: GSM4333241: e11.5_mouse_Hox13KO_HOXA11_ChIPseq_rep1; Mus musculus; ChIP-Seq
library_construction_protocol: HOXA11 ChIP was performed in forelimb buds of CD1 (wild type), Prx1Cre; RosaHoxa11/Hoxa11 (RA11KI) and Hox13-/- mice at E11.5 in the same conditions as previously described for HOX13 ChIP 7. Chromatin was cross-linked using a combination of disuccinimidyl glutarate (DSG) and formaldehyde and sonicated using Fisher Scientific, Model 100 sonic dismembrator to obtain fragments between 100-600 bp. Protein A and Protein G Dynabeads (Invitrogen) were incubated for 6 hours at 4C with 5ug HOXA11 (SAB1304728, Sigma) antibody. The chromatin was coupled to the beads overnight at 4C. The immunoprecipitated samples were then sequentially washed in low salt (1% Triton, 0,1% SDS, 150 mM NaCl, 20 mM Tris (pH8), 2 mM EDTA), high salt (1% Triton, 0,1% SDS, 500 mM NaCl, 20 mM Tris pH8, 2 mM EDTA), LiCl (1% NP-40, 250 mM LiCl, 10 mM Tris (pH8), 1 mM EDTA) and TE buffer (50 mM NaCl, 10 mM Tris (pH8), 1 mM EDTA). The DNA was then purified on QIAquick columns (Qiagen). Library and flow cells were prepared by the IRCM Molecular Biology Core Facility according to Illumina's recommendations and sequenced on Illumina Hiseq 2500 in a 50 cycles paired-end configuration.
identifiers.namespace: GEO
identifiers.id: GSM4333241