GSM5740056: TKS120 Thymus sonic in vitro Satb1 protein low (BUR reference map); Mus musculus; ChIP-Seq
GSM5740056
The procedure is described in Kohwi-Shigematsu, et al.,2012 (Methods, 58, 243-254).  Briefly, pure genomic DNA isolated from mouse thymocytes was sonicated with BioRuptor (UCD-200, Diagenode) to generate DNA fragments between 300 and 600 bp.  The resulting DNA fragments were allowed to bind to his-tagged Satb1recombinant protein. His-Satb1-DNA complexes are captured by incubation with the pre-blocked MagneHis beads for 5 min at 25oC. The beads were  then washed three times with Binding Buffer. His-Satb1-DNA complexes are eluted from the beads, and the DNA fragments were purified. Briefly for library construction, ChIP DNA fragments were end-repaired and ligated to indexed sequencing adapters (NEB or Rubicon library kit).  Using SPRI beads (Beckmancoulter), small fragments (<200bp) were removed from the amplified library. The size-selected library was sequenced to 50bp single-end read lengths on an Illumina HiSeq4000 (Vincent J. Coates Genomic Sequencing Laboratory, UC Berkeley).