GSM7061509: K562 cells, SF3B1 K700E, 4 days, replicate 2 [DRIPseq_K562_SF3B1K700E_Replicate2]; Homo sapiens; OTHER
Digested DNA was purified by phenol/chloroform/isoamyl alcohol extraction, ethanol precipitation and quantified by Nanodrop. 4 μg of digested DNA was incubated with 10 μg of S9.6 antibody overnight at 4 °C on a rotator. DNA/antibody complexes were added for 2 hours at 4 °C to Agarose Protein-A/G beads. Immunoprecipitated DNA was eluted by incubating with an elution buffer containing proteinase K at 55 °C for 45 min on a rotator. The eluent was precipitated by phenol/chloroform/isoamylalcohol extraction and ethanol precipitation. The pulled down material and input DNA were then sonicated, size-selected, and ligated to Illumina barcoded adaptors, using TruSeq ChIP Sample Preparation Kit or ThruPLEX® DNA-seq Kit