GSM7140984: priB_Rif1WT_ChIPseq_rep1; Mus musculus; ChIP-Seq
Chromatin immunoprecipitation was performed as described Foti et al. 2021. In summary, primary B cells were isolated from Rif1FH/FH and C57BL/6 (Rif1WT/WT) mice spleens using anti-CD43 MicroBeads (Miltenyi Biotec) and expanded in complete RPMI containing 5 µg/ml LPS (Sigma-Aldrich) and 5 ng/ml mouse recombinant IL-4 (Sigma-Aldrich) to allow B cell activation and class switch recombination to IgG1. B cells were harvested 72h after activation and 4 x 107 cells were cross-linked by using 2 mM disuccinimidyl glutarate (ThermoFisher 20593) in PBS for 45 min and 1% formaldehyde for 5 min (Thermo Scientific 28908). The reaction was quenched with 0.125 M glycine. Cells were washed thrice with ice-cold PBS and lysed in SDS lysis buffer. Chromatin fragmentation was performed using a Covaris E220 sonicator to obtain fragments between 200 and 600 bp. Chromatin was quantified with a ND-1000 NanoDrop spectrophotometer. Immunoprecipitation was performed with 0.5 μg of anti-HA antibody (Santa Cruz sc-7392) and 50 µl of Dynabeads protein G (Thermofisher 10003D) and 25 μg chromatin ChIP libraries were prepared by NEBNext ultra II DNA library preparation kit (NEB E7645L) and sequenced on one lane of a NovaSeq 6000 (Illumina) machine 4 x 10^7 cells were cross-linked by using 2 mM disuccinimidyl glutarate  in PBS for 45 min and 1% formaldehyde for 5 min . The reaction was quenched with 0.125 M glycine. Cells were washed thrice with ice-cold PBS and lysed in SDS lysis buffer. Chromatin fragmentation was performed using a Covaris E220 sonicator to obtain fragments between 200 and 600 bp. Chromatin was quantified with a ND-1000 NanoDrop spectrophotometer. Immunoprecipitation was performed with 0.5 μg of anti-HA antibody (Santa Cruz sc-7392) and 50 µl of Dynabeads protein G (Thermofisher 10003D) and 25 μg chromatin