GSM5931406: Pbx1 ChIP-seq Replicate 1; Mus musculus; ChIP-Seq
Embryonic hindlimb buds were isolated from E10.5 mouse embryos, obtained from timed matings of wild-type Swiss Webster mice and immediately crosslinked for 10 min in 1% formaldehyde (Electron Microscopy Sciences, #15710). The crosslinked material was sonicated to 200-500 bp fragments with a Diagenode Bioruptor. Each ChIP-seq assay was performed from pooling 60-80 embryonic hindlimbs, by overnight incubation with specific antibodies (5 µg) at 4°C followed by 30 min incubation with Dynabeads protein A (Invitrogen). After washing, IP and input DNA were de-crosslinked and purified using the MicroChIP DiaPure kit (Diagenode, #C03040001). Libraries were constructed using the MicroPlex Library Preparation Kit v2 (Diagenode, C05010012). Libraries were normalized to 10 nM, pooled, and sequenced in an Illumina HiSeq 4000 to generate 50 base pair single-end reads.