GSM5070265: KO_SUMO1_2; Mus musculus; ChIP-Seq
Dissected ovaries were snap-frozen and stored at −80◦C. Frozen ovaries were crushed in liquid nitrogen using a mini-liquid nitrogen cooled mortar (Bel-Art ref H37260-0100). Powdered tissue samples were immediately fixed in PBS/1% formaldehyde at room temperature for 20 min. Chromatin shearing was performed using qSonica Sonicator.  Chromatin immunoprecipitation (IP) with homemade rabbit anti SUMO1 and anti SUMO2 were performed on 20µg of chromatin using Low cell ChIPseq kit (Active motif, ref 10489). For each genotype, two independent ChIPseq libraries were prepared. For Each library three independent immunoprecipitation were pooled, each IP prepared from ovaries of two different animals. Libraries were prepared using Next Gen DNA Library Kit which contains molecular identifiers (MIDs) used to remove PCR duplicates, and Next Gen Indexing kit (Active motif, ref 104895).