GSM8866295: Input E16 OB; Mus musculus; ChIP-Seq
Both OB, BG and Cortices were fixed in 1.5 % formaldehyde and lysed with a hypotonic buffer to obtain the nuclei; these were then lysed in 1% SDS buffer and the chromatin was sheared into 300-1000 bp fragments by sonicating using a bioruptor (Diagenode). Immunoprecipitation (IP) reactions were performed with the sheared chromatin diluted 1/10 times with “dilution buffer” and 5 µg DLX2 specific Ab was added. Negative control ChIP reactions were set-up with a rabbit IgG (5µg). Antibody/chromatin complexes were purified using Dynabeads (Invitrogen) and washed extensively in “wash buffers” (low salt, high salt, LiCl, and TE).Complexes were eluted in 1% SDS, 10mM Sodium bicarbonate buffer at 65˚C for 10 min. Libraries were prepared using an Ovation Ultralow DR Multiplex System (Nugen) and QC tested on a Bioanalyzer (Agilent). The libraries were sequenced as single-ended 50 nucleotide reads on a HiSeq 4000 or NovaSeq  (Illumina) at Center for Advanced Technology (UCSF).