A front limb of a quadruped. (The Random House College Dictionary, 1980)
Attributes by original data submitter
Sample
source_name
forelimb bud
age
e11.5
strain
mixed background C57Bl/6 x 129
genotype
Prrx1Cre;RosaHoxa11/Hoxa11
chip antibody
HOXA11 sigma: sab1304728
tissue
forelimb buds
Sequenced DNA Library
library_strategy
ChIP-Seq
library_source
GENOMIC
library_selection
ChIP
library_construction_protocol
HOXA11 ChIP was performed in forelimb buds of CD1 (wild type) and Prx1Cre; RosaHoxa11/Hoxa11 (RA11KI) 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 4C with 5ug HOXA11 (SAB1304728, Sigma) antibody. The chromatin was coupled to the beads overnight at 4C. 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.