GSM9038497: Ig3_PS_Ca_sti_Rad21_1; Mus musculus; ChIP-Seq
10 million CH12F3 cell lines treated with  anti-CD40+IL4+TGF-β for 72 hrs. were collected and cross-linked with 1% formaldehyde, followed by quenching with glycine. The cross-linked cells were sonicated in sonication buffer (10 mM Tris pH 8.0, 0.25% SDS, 2 mM EDTA and protease inhibitors) to get chromatin fragments with the size between 200bp to 700bp. Equal amount of chromatin was used for immunoprecipitation with specific antibodies including anti-Rad21(abcam, ab992). After incubating the antibodies with sonicated chromatin at 4℃overnight, Protein A magnetic beads (ThermoFisher, 10002D) were added into the chromatin for another 2 hours to enrich antibody-bound chromatin. After 3 times of wash to remove unspecific binding of chromatin to the antibodies, the chromatin on beads was tagmented by Tn5 transponsase and eluted from beads. Eluted ChIP DNA was subjected to constructing libraries using NEBNext High-Fidelity PCR Master Mix (New England Biolabs, M0541) and illumina index primers. The final ChIP-seq libraries were size-selected by SPRIselect beads (Beckman Coulter, B23318) and sequenced via paired-end 150bp sequencing on Novoseq 6000.